Methods of assessing suitability of use of pharmaceutical compositions of albumin and paclitaxel

ABSTRACT

The present invention provides methods of assessing suitability of a pharmaceutical composition for medical use. The pharmaceutical composition comprises nanoparticles comprising paclitaxel coated with albumin and a non-nanoparticle portion comprising albumin and paclitaxel.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/062,050, filed Mar. 5, 2016, which claims priority of U.S.Provisional Application No. 62/129,012, filed Mar. 5, 2015, each ofwhich is incorporated herein by reference in its entirety for allpurposes.

TECHNICAL FIELD

The present invention relates to methods of assessing suitability of useof a pharmaceutical composition of albumin and paclitaxel.

BACKGROUND

Albumin-based pharmaceutical compositions have been developed as a drugdelivery system for delivering substantially water insoluble drugs suchas a taxane. See, for example, U.S. Pat. Nos. 5,916,596, 6,506,405,6,749,868, 6,537,579, 7,820,788, and 7,923,536. ABRAXANE®, analbumin-stabilized nanoparticle formulation of paclitaxel(“nab-paclitaxel”), is a prescription drug approved to treatlife-threatening cancers that affect hundreds of thousands of patientsin the United States. It is indicated for the treatment of metastaticbreast cancer, locally advanced or metastatic non-small cell lung cancer(“NSCLC”), as well as metastatic adenocarcinoma of the pancreas.

It is generally believed that albumin-based nanoparticles, such as thosein nab-paclitaxel sold under the trademark ABRAXANE®, when introducedinto the blood stream, would dissolve into albumin-drug complexes. Suchalbumin-drug complexes utilize the natural properties of albumin totransport and deliver substantially water insoluble drugs to the site ofdisease, such as tumor sites. In addition, the albumin-basednanoparticle technology offers the ability to improve a drug'ssolubility without the need for toxic solvents in the administrationprocess, thus potentially improving safety through the elimination ofsolvent-related side effects.

The disclosures of all publications, patents, patent applications, andpublished patent applications referred to herein are hereby incorporatedherein by reference in their entireties.

BRIEF SUMMARY DESCRIBED HEREIN

The present application in some embodiment provides methods of assessingsuitability of a composition (such as a pharmaceutical composition) formedical use, wherein the composition (such as a pharmaceuticalcomposition) comprises nanoparticles comprising paclitaxel coated withalbumin and a non-nanoparticle portion comprising albumin andpaclitaxel.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin polymers among thealbumin on the nanoparticles, wherein a percentage of albumin polymeramong the albumin on the nanoparticles being about 15% to about 40%(such as about 15% to about 20%, about 20% to about 24.5%, about 24.5%to about 30%, about 30% to about 35%, or about 35% to about 40%) isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thepercentage of albumin monomers among the albumin on the nanoparticles,wherein a percentage of albumin monomers among the albumin on thenanoparticles being about 40% to about 60% (such as about 40% to about55%, about 40% to about 54%, about 40% to about 53%, about 40% to about52%, about 40% to about 50%, about 40% to about 48%, or about 40% toabout 46%) is indicative of suitability of the pharmaceuticalcomposition for medical use.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin monomers among thealbumin on the nanoparticles, wherein a percentage of albumin monomersamong the albumin on the nanoparticles being about 40% to about 60%(such as about 40% to about 55%, about 40% to about 54%, about 40% toabout 53%, about 40% to about 52%, about 40% to about 50%, about 40% toabout 48%, or about 40% to about 46%) is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin monomers among the albumin on the nanoparticlesbeing less than about 52% is indicative of suitability of thepharmaceutical composition for medical use.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin polymers and oligomersamong the albumin on the nanoparticles, wherein a percentage of albuminpolymers and oligomers among the albumin on the nanoparticles being morethan about 35% is indicative of suitability of the pharmaceuticalcomposition for medical use.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin polymers and monomersamong the albumin on the nanoparticles, wherein a percentage of albuminpolymers among the albumin on the nanoparticles being more than about11% and a percentage of albumin monomers among the albumin on thenanoparticles being less than about 54% is indicative of suitability ofthe pharmaceutical composition for medical use.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin polymers and monomersamong the albumin on the nanoparticles, wherein a percentage of albuminpolymers among the albumin on the nanoparticles being more than about18% and a percentage of albumin monomers among the albumin on thenanoparticles being less than about 55% is indicative of suitability ofthe pharmaceutical composition for medical use.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin polymers, oligomers,and monomers among the albumin on the nanoparticles, wherein the ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 62% indicative of suitability of the pharmaceuticalcomposition for medical use.

In some embodiments according to any of the methods described above, themethod further comprises determining the weight percentage of thealbumin in the nanoparticles, wherein a weight percentage of the albuminin the nanoparticles being about 15% to about 30% (such as about 20% toabout 25%, about 15% to about 24%, or about 15% to about 20%) isindicative of suitability of the pharmaceutical composition for medicaluse.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the weight percentage of the albumin in thenanoparticles, wherein a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%) is indicative ofsuitability of the pharmaceutical composition for medical use.

In some embodiments according to any of the methods described above, themethod further comprises determining the weight ratio of albumin topaclitaxel in the nanoparticles, wherein an albumin to paclitaxel ratioof about 1:2 to about 1:6 in the nanoparticles is indicative ofsuitability of the pharmaceutical composition for medical use.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the weight ratio of albumin to paclitaxel in thenanoparticles, wherein an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles is indicative of suitability of thepharmaceutical composition for medical use.

In some embodiments according to any of the methods described above, themethod further comprises determining the morphology of the nanoparticlesunder cryo-TEM, wherein an irregular shape of the nanoparticles isindicative of suitability of the pharmaceutical composition for medicaluse.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the morphology of the nanoparticles undercryo-TEM, wherein an irregular shape of the nanoparticles is indicativeof suitability of the pharmaceutical composition for medical use.

In some embodiments according to any of the methods described above, themethod further comprises determining the thickness of the albumincoating of the nanoparticles under cryo-TEM, wherein a thickness ofabout 5-7 nm (such as about 6 nm) is indicative of suitability of thepharmaceutical composition for medical use.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the thickness of the albumin coating of thenanoparticles under cryo-TEM, wherein a thickness of about 5-7 nm (suchas about 6 nm) is indicative of suitability of the pharmaceuticalcomposition for medical use.

In some embodiments according to any of the methods described above,there is provided a method of assessing suitability of a pharmaceuticalcomposition for medical use in a human individual, wherein thepharmaceutical composition comprises nanoparticles comprising paclitaxelcoated with albumin and a non-nanoparticle portion comprising albuminand paclitaxel, the method comprising: determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue; wherein an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use.

In some embodiments according to any of the methods described above, themethod further comprises determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor; wherein an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse.

In some embodiments according to any of the methods described above, themethod further comprises determining the solubility of thepharmaceutical composition, wherein a solubility of about 50 μg/ml toabout 80 μg/ml in a 5% human albumin solution is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the determination of solubility is carried out afterstorage.

In some embodiments according to any of the methods described above, themethod further comprises determining the paclitaxel crystallinity of thepharmaceutical composition, wherein a non-crystalline state of thepaclitaxel is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the determination ofpaclitaxel crystalline state is carried out after storage. In someembodiments, the paclitaxel crystallinity is determined by X-raydiffraction, polarized light microscopy, or both.

In some embodiments according to any of the methods described above, themethod further comprises determining the paclitaxel recovery following a0.2 micron filtration of the pharmaceutical composition, wherein apaclitaxel recovery of at least about 80% is indicative of suitabilityof the pharmaceutical composition for medical use. In some embodiments,the determination of paclitaxel recovery is carried out after storage.

One aspect of the present application provides a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the solubility, paclitaxel crystallinity, and apaclitaxel recovery following a 0.2 micron filtration of thepharmaceutical composition, wherein a solubility of about 50 μg/ml toabout 80 μg/ml in a 5% human albumin solution, a non-crystalline stateof the paclitaxel, and a paclitaxel recovery date of at least about 80%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, the method is carried out afterstorage.

In some embodiments according to any of the methods described above, themethod further comprises determining the binding affinity of albumin topaclitaxel in the pharmaceutical composition. In some embodiments, thebinding affinity is determined by equilibrium dialysis, FTIR, NMR, or acombination thereof.

In some embodiments according to any of the methods described above, themethod further comprises determining the surface-to-volume ratio of thenanoparticles in the pharmaceutical composition.

In some embodiments according to any of the methods described above, themethod further comprises determining the percentage of albumin dimersamong the albumin on the nanoparticles, wherein a percentage of about15% to about 30% of albumin dimers among the albumin on thenanoparticles is indicative of the pharmaceutical composition formedical use.

In some embodiments according to any of the methods described above, themethod further comprises determining the percentage of albumin oligomersamong the albumin on the nanoparticles, wherein a percentage of about 7%to about 15% of albumin oligomers among the albumin on the nanoparticlesis indicative of the pharmaceutical composition for medical use.

In some embodiments according to any of the methods described above, themethod further comprises determining the percentage of albumin monomers,dimers, oligomers, or polymers among the total albumin in thepharmaceutical composition. In some embodiments, the percentage ofalbumin monomers, dimers, oligomers, or polymers is carried out bysize-exclusion chromatography.

In some embodiments according to any of the methods described above, themethod further comprises determining the particle size of thenanoparticles. In some embodiments, the particle size of thenanoparticles is determined by dynamic light scattering.

In some embodiments according to any of the methods described above, themethod further comprises determining the polydispersity index of thenanoparticles in the pharmaceutical composition.

In some embodiments according to any of the methods described above, themethod further comprises determining the span of size distribution((Dv₉₀−Dv₁₀)/Dv₅₀) of the nanoparticles in the pharmaceuticalcomposition. Dv₅₀ refers to the volume-weighted median particlediameter. Dv₉₀ refers to the particle diameter where 90% of the volumeof all nanoparticles is contained in nanoparticles with smallerdiameters. Dv₁₀ refers to the particle diameter where 10% of the volumeof all nanoparticles is contained in nanoparticles with smallerdiameters.

In some embodiments according to any of the methods described above, themethod further comprises determining the surface potential of thenanoparticles.

In some embodiments according to any of the methods described above, themethod further comprises determining the percentage of the paclitaxel inthe nanoparticles among the total paclitaxel in the pharmaceuticalcomposition. In some embodiments, the percentage of the paclitaxel inthe nanoparticles is determined by reversed-phase HPLC.

In some embodiments according to any of the methods described above, themethod further comprises determining the percentage of the albumin thatis in the non-nanoparticle portion among the total albumin in thepharmaceutical composition. In some embodiments, the percentage of thealbumin is determined by size-exclusion chromatography.

In some embodiments according to any of the methods described above, themethod further comprises determining the stability of the pharmaceuticalcomposition. In some embodiments, the stability is determined afterstorage.

In some embodiments according to any of the methods described above, themethod further comprises determining tumor distribution of paclitaxelupon administration in vivo.

In some embodiments, the method comprises determining tumor distributionof paclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue.

In some embodiments according to any of the methods described above, theweight ratio of the total albumin to the total paclitaxel in thepharmaceutical composition is about 3:1 to about 7.9:1 or about 10:1 toabout 17:1.

In some embodiments according to any of the methods described above, thealbumin is human albumin.

In some embodiments according to any of the methods described above, theaverage particle size of the nanoparticles is less than about 200 nm(such as about 120 nm to about 140 nm, for example about 130 nm).

In a further aspect of the present application, there is provided amethod of validating a commercial batch of a pharmaceutical compositionfor medical use in a human individual, wherein the pharmaceuticalcomposition comprises nanoparticles comprising paclitaxel coated withalbumin and a non-nanoparticle portion comprising albumin andpaclitaxel, and wherein the method comprises 1) obtaining a sample fromthe commercial batch, and 2) assessing suitability of the sample formedical use according to any one of the methods of assessing asdescribed above.

In a further aspect of the present application, there is provided acommercial batch of a pharmaceutical composition for medical use in ahuman individual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, and whereinthe commercial batch is validated by assessment of suitability formedical use according to any one of the methods of assessing asdescribed above.

Also provided are kits, medicines, and articles of manufacturecomprising any one of the compositions (such as pharmaceuticalcompositions) described above.

These and other aspects and advantages of the present invention willbecome apparent from the subsequent detailed description and theappended claims. It is to be understood that one, some, or all of theproperties of the various embodiments described herein may be combinedto form other embodiments of the present invention.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A-1C show representative imaging of pancreatic MIA PaCa-2xenograft tumors 24 hours (FIG. 1A), 48 hours (FIG. 1B), and 72 hours(FIG. 1C) post-injection with the nab-paclitaxel sold under thetrademark ABRAXANE® (ABX). Mitotically-arrested cells were stained withan anti-pHH3 antibody (white).

FIGS. 1D-1F show representative imaging of pancreatic MIA PaCa-2xenograft tumors 24 hours (FIG. 1D), 48 hours (FIG. 1E), and 72 hours(FIG. 1F) post-injection with a DMSO formulation of paclitaxel(PTX:DMSO). Mitotically-arrested cells were stained with an anti-pHH3antibody (white).

FIGS. 1G-1I show representative imaging of pancreatic MIA PaCa-2xenograft tumors 24 hours (FIG. 1G), 48 hours (FIG. 1H), and 72 hours(FIG. 1I) post-injection with a Cremophor EL formulation of paclitaxel(PTX:CrEL). Mitotically-arrested cells were stained with an anti-pHH3antibody (white).

FIGS. 2A-2C show the fraction of pHH3 positive cells (fraction pHH3+)versus the radial distance (μm) as measured from the injection site forpancreatic MIA PaCa-2 xenograft tumors 24 hours (FIG. 2A), 48 hours(FIG. 2B), and 72 hours (FIG. 2C) post-injection with either ABX,PTX:DMSO, or PTX:CrEL.

FIG. 3A shows the fraction pHH3+ versus the radial distance (μm) asmeasured from the injection site for A2058 tumor xenografts 24 hourspost-injection with either ABX, PTX:CrEL, CrEL, or PBS.

FIG. 3B shows the fraction pHH3+ versus the radial distance (μm) asmeasured from the injection site for H2122 tumor xenografts 24 hourspost-injection with either ABX, PTX:DMSO, DMSO, or PBS.

FIGS. 4A-4C show the fraction pHH3+ versus the radial distance (μm) asmeasured from the injection site for pancreatic MIA PaCa-2 xenografttumors at 24 hours post-injection with either 1.6 mg/mL ABX, 1.6 mg/mLPTX:DMSO, or 1.6 mg/mL PTX:CrEL (FIG. 4A); either 2.5 mg/mL ABX, 2.5mg/mL PTX:DMSO, or 2.5 mg/mL PTX:CrEL (FIG. 4B); and either 4.75 mg/mLABX, 4.75 mg/mL PTX:DMSO, or 4.75 mg/mL PTX:CrEL (FIG. 4C). The level ofbackground signal is indicated with a dashed line.

FIG. 5 shows a chromatogram from the separation of polymeric,oligomeric, dimeric, and monomeric albumin on nanoparticles from apharmaceutical composition using size-exclusion chromatography.

FIG. 6 shows a diagram of a UV-Vis spectrophotometer optical system.

FIG. 7 shows in vitro dissolution kinetics of the nab-paclitaxel soldunder the trademark ABRAXANE® in water at 100 μg/ml paclitaxelconcentration, as measured at 340 nm by a UV-Vis spectrophotometer witha 295 nm low wavelength cut-off filter.

FIG. 8 shows a bar graph of the albumin as a percentage of thenanoparticle mass.

FIG. 9 shows a bar graph of the percentage of albumin on thenanoparticles in the form of monomers.

FIG. 10 shows a bar graph of the percentage of albumin on thenanoparticles in the form of dimers.

FIG. 11 shows a bar graph of the percentage of albumin on thenanoparticles in the form of oligomers.

FIG. 12 shows a bar graph of the percentage of albumin on thenanoparticles in the form of polymers.

FIG. 13 shows a bar graph of the percentage of albumin on thenanoparticles in the form of monomers (M) and dimers (D).

FIG. 14 shows a bar graph of the percentage of albumin on thenanoparticles in the form of monomers (M) minus the percentage ofalbumin on the nanoparticles in the form of dimers (D).

FIG. 15 shows a bar graph of the percentage of albumin on thenanoparticles in the form of monomers (M) and oligomers (O).

FIG. 16 shows a bar graph of the percentage of albumin on thenanoparticles in the form of monomers (M) and polymers (P).

FIG. 17 shows a bar graph of the percentage of albumin on thenanoparticles in the form of monomers (M) minus the percentage ofalbumin on the nanoparticles in the form of polymers (P).

FIG. 18 shows a bar graph of the percentage of albumin on thenanoparticles in the form of dimers (D) and oligomers (O).

FIG. 19 shows a bar graph of the percentage of albumin on thenanoparticles in the form of dimers (D) and polymers (P).

FIG. 20 shows a bar graph of the percentage of albumin on thenanoparticles in the form of oligomers (O) and polymers (P).

FIG. 21 shows a bar graph of the ratio (reported as a percentage) of thepercentage of albumin on the nanoparticles in the form of dimers (D)divided by the percentage of albumin on the nanoparticles in the form ofmonomers (M).

FIG. 22 shows a bar graph of the ratio (reported as a percentage) of thepercentage of albumin on the nanoparticles in the form of oligomers (O)divided by the percentage of albumin on the nanoparticles in the form ofmonomers (M).

FIG. 23 shows a bar graph of the ratio (reported as a percentage) of thepercentage of albumin on the nanoparticles in the form of polymers (P)divided by the percentage of albumin on the nanoparticles in the form ofmonomers (M).

FIG. 24 shows a bar graph of the ratio (reported as a percentage) of thepercentage of albumin on the nanoparticles in the form of polymers (P)and oligomers (O) divided by the percentage of albumin on thenanoparticles in the form of monomers (M).

FIG. 25 shows a bar graph of the ratio (reported as a percentage) of thepercentage of albumin on the nanoparticles in the form of polymers (P)and oligomers (O) divided by the percentage of albumin on thenanoparticles in the form of monomers (M) minus dimers (D).

DETAILED DESCRIPTION

The present application provides methods of assessing suitability formedical use (for example, medical use in a human individual) of analbumin-based nanoparticle composition (for example a pharmaceuticalcomposition) by determining one or a number of physicochemicalcharacteristics and functional attributes of the composition. Thepharmaceutical compositions comprise: a) nanoparticles comprisingpaclitaxel coated with albumin, and b) a non-nanoparticle portioncomprising albumin and paclitaxel. The methods comprise determination ofat least one (such as at least any of 2, 3, 4, 5, 6, 7, or 8) of thefollowing characteristics or attributes: i) the oligomeric status of thealbumin on the nanoparticles, including percentage of albumin polymersand/or monomers on the nanoparticles; ii) the percent by weight of thealbumin in the nanoparticles; iii) the weight ratio of the albumin tothe paclitaxel in the nanoparticles; iv) particle morphology, includingshape, thickness of the coating, or surface-to-volume ratio; v)distribution of paclitaxel in a tumor tissue upon administration of thecomposition; vi) particle solubility; vii) paclitaxel crystallinity; andviii) paclitaxel recovery following a 0.2 micron filtration. The methodsmay further comprise determination of at least one (such as at least anyof 2, 3, 4, 5, 6, 7, 8, or 9) of the following characteristics orattributes: 1) binding affinity of albumin to paclitaxel in thecomposition (for example by equilibrium dialysis, FTIR, NMR, or acombination thereof); 2) surface-to-volume ratio; 3) percentage ofalbumin dimers and/or oligomers among the albumin on the nanoparticles;4) distribution of the total paclitaxel and/or the total albumin betweenthe nanoparticles and the non-nanoparticle portion; 5) oligomeric statusof the total albumin in the composition; 6) particle size of thenanoparticles, including average particle size, polydispersity, and/orsize distribution; 7) surface potential; 8) in vitro release kinetics;and 9) physical stability.

The methods provided herein are useful, for example, for validatingand/or releasing a commercial batch of an albumin-based paclitaxelnanoparticle composition.

The compositions (such as pharmaceutical compositions) described herein,once determined to be suitable for medical use in a human individual,can be useful for treating various diseases, such as cancer. The presentapplication thus also provides compositions (such as pharmaceuticalcompositions, including for example commercial batches) determined to besuitable for medical use, as well as methods of using such compositions(such as pharmaceutical compositions) for the treatment of diseases,including cancer. Also provided herein are kits, medicines, and dosageforms comprising the compositions (such as pharmaceutical compositions)described herein and for use in methods described herein.

The exemplary embodiments provided herein disclose pharmaceuticalcompositions. It is to be understood that these are exemplarycompositions and that these descriptions apply equally to and describeother compositions of the invention as provided herein, such ascompositions having any of the characteristics defined in theseexemplary embodiments.

Definitions

The term “individual” refers to a mammal and includes, but is notlimited to, human, bovine, horse, feline, canine, rodent, or primate.

It is understood that aspects and embodiments described herein include“consisting” and/or “consisting essentially of” aspects and embodiments.

Reference to “about” a value or parameter herein includes (anddescribes) variations that are directed to that value or parameter perse. For example, description referring to “about X” includes descriptionof “X.”

The term “about X-Y” used herein has the same meaning as “about X toabout Y.”

As used herein and in the appended claims, the singular forms “a,” “or,”and “the” include plural referents unless the context clearly dictatesotherwise.

“Monomers” used herein refers to a single albumin molecule withoutintermolecular disulfide bonds.

“RRT” used herein refers to the retention time relative to the albuminmonomers retention time on a size-exclusion HPLC chromatography.

“Dimers” used herein refers to albumin species having an RRT of about0.86 to about 0.97.

“Oligomers” used herein refers to albumin species having an RRT of about0.70 to about 0.85.

“Polymers” used herein refers to albumin species having an RRT of about0.57 to about 0.69.

“The total albumin” in a composition (such as a pharmaceuticalcomposition) comprises the albumin on the nanoparticles and the albuminin the non-nanoparticle portion of the composition. “The albumin on thenanoparticles” or “the albumin in the nanoparticles” refers to thealbumin coated on the paclitaxel in the nanoparticles, or the albumincoating of the nanoparticles. “The total paclitaxel” in a composition(such as a pharmaceutical composition) comprises the paclitaxel in thenanoparticles and the paclitaxel in the non-nanoparticle portion of thecomposition.

“Weight percentage of albumin in the nanoparticles” used herein refersto the weight percentage of albumin in the total weight of thenanoparticles.

“Weight ratio of albumin to paclitaxel in the nanoparticles” used hereinrefers to the weight ratio of albumin on the nanoparticles to thepaclitaxel on the nanoparticles.

Methods of Assessing Suitability of Albumin-Based PaclitaxelNanoparticle Compositions for Medical Use

The present application provides a method of assessing suitability of acomposition (also referred to as “albumin-based paclitaxel nanoparticlecomposition”) for medical use in an individual, wherein the compositioncomprises nanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel. The methodscomprise determination of at least one (such as at least any of 2, 3, 4,5, 6, 7, or 8) of the following characteristics or attributes: i) theoligomeric status of the albumin on the nanoparticles (i.e. the albumincoating), including percentage of albumin polymers and/or monomers inthe nanoparticles; ii) the percent by weight of the albumin in thenanoparticles; iii) the weight ratio of the albumin to the paclitaxel inthe nanoparticles; iv) particle morphology, including shape, thicknessof the coating, and surface-to-volume ratio; v) distribution ofpaclitaxel in a tumor tissue upon administration of the composition (forexample upon direct injection of the composition directly into the tumortissue); vi) particle solubility; vii) paclitaxel crystallinity; andviii) paclitaxel recovery following a 0.2 micron filtration. The methodsmay further comprise determination of at least one (such as at least anyof 2, 3, 4, 5, 6, 7, 8, 9, or 10) of the following characteristics orattributes: 1) binding affinity of albumin to paclitaxel in thecomposition (for example by equilibrium dialysis, FTIR, NMR, or acombination thereof); 2) surface-to-volume ratio; 3) percentage ofalbumin dimers and/or oligomers among the albumin on the nanoparticles;4) distribution of the total paclitaxel and/or the total albumin betweenthe nanoparticles and the non-nanoparticle portion; 5) oligomeric statusof the total albumin in the composition; 6) particle size of thenanoparticles, including average particle size, polydispersity, and/orsize distribution; 7) surface potential; 8) in vitro release kinetics;9) physical stability; and, in some embodiments, 10) paclitaxel tumordistribution in vivo.

Unless otherwise indicated, discussion of a certain parameter as beingindicative of suitability for medical use suggest that such parametermay be determined in the method described herein. The method thus, insome embodiments, encompasses a step of determining such a parameter.

The compositions (such as pharmaceutical compositions) described hereincan be in liquid or powder forms. For example, in some embodiments, thecomposition is a liquid nanoparticle suspension (for example prior tolyophilization). In some embodiments, the composition is a reconstitutedsuspension (e.g., in an aqueous solution such as a saline solution). Insome embodiments, the paclitaxel concentration in the suspension isabout any of 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6 mg/ml, 7 mg/ml, 8mg/ml, 9 mg/ml, or 10 mg/ml. In some embodiments, the paclitaxel in thesuspension is about 5 mg/ml. In some embodiments, the composition islyophilized. In some embodiments, the composition is sterile. In someembodiments, the composition is contained in a sealed vial.

Thus, in some embodiments, there is provided a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin polymers among thealbumin on the nanoparticles, wherein a percentage of albumin polymeramong the albumin on the nanoparticles being about 15% to about 40%(such as about 15% to about 20%, about 20% to about 24.5%, about 24.5%to about 30%, about 30% to about 35%, or about 35% to about 40%) isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the size ofthe nanoparticles (for example by dynamic light scattering). In someembodiments, the method further comprises determining the polydispersityindex of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the span of sizedistribution ((Dv₉₀-Dv₁₀)/Dv₅₀) of the nanoparticles in thepharmaceutical composition. In some embodiments, the method furthercomprises determining the surface potential of the nanoparticles. Insome embodiments, the method further comprises determining thepercentage of the paclitaxel in the nanoparticles among the totalpaclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀-Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, wherein a percentage of albumin polymer among the albuminon the nanoparticles being about 15% to about 40% (such as about 15% toabout 20%, about 20% to about 24.5%, about 24.5% to about 30%, about 30%to about 35%, or about 35% to about 40%) (such as any of about 15% toabout 20%, about 20% to about 24.5%, about 24.5% to about 30%, about 30%to about 35%, or about 35% to about 40%), and a percentage of albuminmonomer among the albumin on the nanoparticles being at least about 40%to about 60% (such as about 40% to about 55%, about 40% to about 54%,about 40% to about 53%, about 40% to about 52%, about 40% to about 50%,about 40% to about 48%, or about 40% to about 46%) is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the method further comprises determining the solubility ofthe pharmaceutical composition (including determining solubility afterstorage). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀-Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, wherein aweight percentage of the albumin in the nanoparticles being about 15% toabout 30% (such as about 20% to about 25%, about 15% to about 24%, orabout 15% to about 20%) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles and determining the weight percentage of the albumin inthe nanoparticles, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%) and a weightpercentage of the albumin in the nanoparticles being about 15% to about30% (such as about 20% to about 25%, about 15% to about 24%, or about15% to about 20%) is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thepaclitaxel crystallinity of the pharmaceutical composition (for exampleby X-ray diffraction and/or polarized light microscopy, includingdetermining crystallinity after storage). In some embodiments, themethod further comprises determining the paclitaxel recovery following a0.2 micron filtration (including determining recovery after storage). Insome embodiments, the method further comprises determining bindingaffinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀-Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles and determining the weight percentage of the albumin inthe nanoparticles, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%) and a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%) is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀-Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles and determining the weight percentage of the albumin inthe nanoparticles, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), and aweight percentage of the albumin in the nanoparticles being about 15% toabout 30% (such as about 20% to about 25%, about 15% to about 24%, orabout 15% to about 20%) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight ratio of albumin to paclitaxel in the nanoparticles, whereinan albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin monomer among the albumin on the nanoparticles being less thanabout 51% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersand oligomers among the albumin on the nanoparticles being more thanabout 35% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersamong the albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles and determining the weight ratio of albumin to paclitaxelin the nanoparticles, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%) and an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles and determining the weight ratio of albumin to paclitaxelin the nanoparticles, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%), and an albumin to paclitaxel ratio of about 1:2to about 1:6 in the nanoparticles is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles and determining the weight ratio of albumin to paclitaxelin the nanoparticles, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), and a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), and analbumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin monomer among the albumin on the nanoparticles being less thanabout 51% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersand oligomers among the albumin on the nanoparticles being more thanabout 35% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersamong the albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, anddetermining the weight ratio of albumin to paclitaxel in thenanoparticles, wherein a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%) and an albuminto paclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the weight ratio of albumin to paclitaxelin the nanoparticles, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%) and an albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin monomer among the albumin on the nanoparticles being less thanabout 51% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersand oligomers among the albumin on the nanoparticles being more thanabout 35% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersamong the albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the weight ratio of albumin to paclitaxelin the nanoparticles, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%) and an albuminto paclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the weight ratio of albumin to paclitaxelin the nanoparticles, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), a weightpercentage of the albumin in the nanoparticles being about 15% to about30% (such as about 20% to about 25%, about 15% to about 24%, or about15% to about 20%), and an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition).

In some embodiments, the method further comprises determining thestability of the pharmaceutical composition (including determiningstability after storage). In some embodiments, the method furthercomprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe morphology of the nanoparticles under cryo-TEM, wherein an irregularshape of the nanoparticles is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles and determining the morphology of the nanoparticles undercryo-TEM, wherein a percentage of albumin polymer among the albumin onthe nanoparticles being about 15% to about 40% (such as about 15% toabout 20%, about 20% to about 24.5%, about 24.5% to about 30%, about 30%to about 35%, or about 35% to about 40%) and an irregular shape of thenanoparticles is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thepaclitaxel crystallinity of the pharmaceutical composition (for exampleby X-ray diffraction and/or polarized light microscopy, includingdetermining crystallinity after storage). In some embodiments, themethod further comprises determining the paclitaxel recovery following a0.2 micron filtration (including determining recovery after storage). Insome embodiments, the method further comprises determining bindingaffinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles and determining the morphology of the nanoparticles undercryo-TEM, wherein a percentage of albumin monomers among the albumin onthe nanoparticles being about 40% to about 60% (such as about 40% toabout 55%, about 40% to about 54%, about 40% to about 53%, about 40% toabout 52%, about 40% to about 50%, about 40% to about 48%, or about 40%to about 46%) and about 40% to about 60% (such as about 40% to about55%, about 40% to about 54%, about 40% to about 53%, about 40% to about52%, about 40% to about 50%, about 40% to about 48%, or about 40% toabout 46%) and an irregular shape of the nanoparticles is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the method further comprises determining the solubility ofthe pharmaceutical composition (including determining solubility afterstorage). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles and determining the morphology of the nanoparticles undercryo-TEM, wherein a percentage of albumin polymer among the albumin onthe nanoparticles being about 15% to about 40% (such as about 15% toabout 20%, about 20% to about 24.5%, about 24.5% to about 30%, about 30%to about 35%, or about 35% to about 40%), a percentage of albuminmonomer among the albumin on the nanoparticles being at least about 40%to about 60% (such as about 40% to about 55%, about 40% to about 54%,about 40% to about 53%, about 40% to about 52%, about 40% to about 50%,about 40% to about 48%, or about 40% to about 46%), and an irregularshape of the nanoparticles is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, anddetermining the morphology of the nanoparticles under cryo-TEM, whereina weight percentage of the albumin in the nanoparticles being about 15%to about 30% (such as about 20% to about 25%, about 15% to about 24%, orabout 15% to about 20%) and an irregular shape of the nanoparticles isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the morphology of the nanoparticles undercryo-TEM, wherein a percentage of albumin polymer among the albumin onthe nanoparticles being about 15% to about 40% (such as about 15% toabout 20%, about 20% to about 24.5%, about 24.5% to about 30%, about 30%to about 35%, or about 35% to about 40%), a weight percentage of thealbumin in the nanoparticles being about 15% to about 30% (such as about20% to about 25%, about 15% to about 24%, or about 15% to about 20%),and an irregular shape of the nanoparticles is indicative of suitabilityof the pharmaceutical composition for medical use. In some embodiments,a percentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the morphology of the nanoparticles undercryo-TEM, wherein a percentage of albumin monomers among the albumin onthe nanoparticles being about 40% to about 60% (such as about 40% toabout 55%, about 40% to about 54%, about 40% to about 53%, about 40% toabout 52%, about 40% to about 50%, about 40% to about 48%, or about 40%to about 46%), a weight percentage of the albumin in the nanoparticlesbeing about 15% to about 30% (such as about 20% to about 25%, about 15%to about 24%, or about 15% to about 20%), and an irregular shape of thenanoparticles is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin monomer among the albumin on the nanoparticles being less thanabout 51% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersand oligomers among the albumin on the nanoparticles being more thanabout 35% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersamong the albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the morphology of the nanoparticles undercryo-TEM, wherein a percentage of albumin polymer among the albumin onthe nanoparticles being about 15% to about 40% (such as about 15% toabout 20%, about 20% to about 24.5%, about 24.5% to about 30%, about 30%to about 35%, or about 35% to about 40%), a percentage of albuminmonomer among the albumin on the nanoparticles being at least about 40%to about 60% (such as about 40% to about 55%, about 40% to about 54%,about 40% to about 53%, about 40% to about 52%, about 40% to about 50%,about 40% to about 48%, or about 40% to about 46%), a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%), and an irregular shape of the nanoparticles is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the method further comprises determining the solubility ofthe pharmaceutical composition (including determining solubility afterstorage). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight ratio of albumin to paclitaxel in the nanoparticles anddetermining the morphology of the nanoparticles under cryo-TEM, whereinan albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles and an irregular shape of the nanoparticles is indicativeof suitability of the pharmaceutical composition for medical use. Insome embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesunder cryo-TEM, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, and anirregular shape of the nanoparticles is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesunder cryo-TEM, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%), an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles and an irregular shape of thenanoparticles is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin monomer among the albumin on the nanoparticles being less thanabout 51% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersand oligomers among the albumin on the nanoparticles being more thanabout 35% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersamong the albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesunder cryo-TEM, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), analbumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, and an irregular shape of the nanoparticles is indicativeof suitability of the pharmaceutical composition for medical use. Insome embodiments, a percentage of albumin monomer among the albumin onthe nanoparticles being less than about 51% is indicative of suitabilityof the pharmaceutical composition for medical use. In some embodiments,a percentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe weight ratio of albumin to paclitaxel in the nanoparticles, anddetermining the morphology of the nanoparticles under cryo-TEM, whereina weight percentage of the albumin in the nanoparticles being about 15%to about 30% (such as about 20% to about 25%, about 15% to about 24%, orabout 15% to about 20%), an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles, and an irregular shape of thenanoparticles is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesunder cryo-TEM, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%), an albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, and an irregular shape of the nanoparticles is indicativeof suitability of the pharmaceutical composition for medical use. Insome embodiments, a percentage of albumin monomer among the albumin onthe nanoparticles being less than about 51% is indicative of suitabilityof the pharmaceutical composition for medical use. In some embodiments,a percentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesunder cryo-TEM, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, and anirregular shape of the nanoparticles is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesunder cryo-TEM, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), a weightpercentage of the albumin in the nanoparticles being about 15% to about30% (such as about 20% to about 25%, about 15% to about 24%, or about15% to about 20%), an albumin to paclitaxel ratio of about 1:2 to about1:6 in the nanoparticles, and an irregular shape of the nanoparticles isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe thickness of the albumin coating of the nanoparticles undercryo-TEM, wherein a thickness of about 5-7 nm (such as about 6 nm) isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles and determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, wherein a percentage of albuminpolymer among the albumin on the nanoparticles being about 15% to about40% (such as about 15% to about 20%, about 20% to about 24.5%, about24.5% to about 30%, about 30% to about 35%, or about 35% to about 40%)and a thickness of about 5-7 nm (such as about 6 nm) is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles and determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, wherein a percentage of albuminmonomers among the albumin on the nanoparticles being about 40% to about60% (such as about 40% to about 55%, about 40% to about 54%, about 40%to about 53%, about 40% to about 52%, about 40% to about 50%, about 40%to about 48%, or about 40% to about 46%) and a thickness of about 5-7 nm(such as about 6 nm) is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin monomer among the albumin on the nanoparticles being less thanabout 51% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersand oligomers among the albumin on the nanoparticles being more thanabout 35% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersamong the albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles and determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, wherein a percentage of albuminpolymer among the albumin on the nanoparticles being about 15% to about40% (such as about 15% to about 20%, about 20% to about 24.5%, about24.5% to about 30%, about 30% to about 35%, or about 35% to about 40%),a percentage of albumin monomer among the albumin on the nanoparticlesbeing at least about 40% to about 60% (such as about 40% to about 55%,about 40% to about 54%, about 40% to about 53%, about 40% to about 52%,about 40% to about 50%, about 40% to about 48%, or about 40% to about46%), and a thickness of about 5-7 nm (such as about 6 nm) is indicativeof suitability of the pharmaceutical composition for medical use. Insome embodiments, a percentage of albumin monomer among the albumin onthe nanoparticles being less than about 51% is indicative of suitabilityof the pharmaceutical composition for medical use. In some embodiments,a percentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, anddetermining the thickness of the albumin coating of the nanoparticlesunder cryo-TEM, wherein a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%) and a thicknessof about 5-7 nm (such as about 6 nm) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀-Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, wherein a percentage of albuminpolymer among the albumin on the nanoparticles being about 15% to about40% (such as about 15% to about 20%, about 20% to about 24.5%, about24.5% to about 30%, about 30% to about 35%, or about 35% to about 40%),a weight percentage of the albumin in the nanoparticles being about 15%to about 30% (such as about 20% to about 25%, about 15% to about 24%, orabout 15% to about 20%), and a thickness of about 5-7 nm (such as about6 nm) is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin monomer amongthe albumin on the nanoparticles being less than about 51% is indicativeof suitability of the pharmaceutical composition for medical use. Insome embodiments, a percentage of albumin polymers and oligomers amongthe albumin on the nanoparticles being more than about 35% is indicativeof suitability of the pharmaceutical composition for medical use. Insome embodiments, a percentage of albumin polymers among the albumin onthe nanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, wherein a percentage of albuminmonomers among the albumin on the nanoparticles being about 40% to about60% (such as about 40% to about 55%, about 40% to about 54%, about 40%to about 53%, about 40% to about 52%, about 40% to about 50%, about 40%to about 48%, or about 40% to about 46%), a weight percentage of thealbumin in the nanoparticles being about 15% to about 30% (such as about20% to about 25%, about 15% to about 24%, or about 15% to about 20%),and a thickness of about 5-7 nm (such as about 6 nm) is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, wherein a percentage of albuminpolymer among the albumin on the nanoparticles being about 15% to about40% (such as about 15% to about 20%, about 20% to about 24.5%, about24.5% to about 30%, about 30% to about 35%, or about 35% to about 40%),a percentage of albumin monomer among the albumin on the nanoparticlesbeing at least about 40% to about 60% (such as about 40% to about 55%,about 40% to about 54%, about 40% to about 53%, about 40% to about 52%,about 40% to about 50%, about 40% to about 48%, or about 40% to about46%), a weight percentage of the albumin in the nanoparticles beingabout 15% to about 30% (such as about 20% to about 25%, about 15% toabout 24%, or about 15% to about 20%), and a thickness of about 5-7 nm(such as about 6 nm) is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin monomer among the albumin on the nanoparticles being less thanabout 51% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersand oligomers among the albumin on the nanoparticles being more thanabout 35% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersamong the albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles.

In some embodiments, the method further comprises determining thepercentage of albumin oligomers among the albumin on the nanoparticles.In some embodiments, the method further comprises determining thepercentage of albumin monomers, dimers, oligomers, or polymers among thetotal albumin in the pharmaceutical composition. In some embodiments,the method further comprises determining the particle size of thenanoparticles (for example by dynamic light scattering). In someembodiments, the method further comprises determining the polydispersityindex of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the span of sizedistribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of the nanoparticles in thepharmaceutical composition. In some embodiments, the method furthercomprises determining the surface potential of the nanoparticles. Insome embodiments, the method further comprises determining thepercentage of the paclitaxel in the nanoparticles among the totalpaclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight ratio of albumin to paclitaxel in the nanoparticles, anddetermining the thickness of the albumin coating of the nanoparticlesunder cryo-TEM, wherein an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles, and a thickness of about 5-7 nm (such asabout 6 nm) is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the thickness of the albumin coatingof the nanoparticles under cryo-TEM, wherein a percentage of albuminpolymer among the albumin on the nanoparticles being about 15% to about40% (such as about 15% to about 20%, about 20% to about 24.5%, about24.5% to about 30%, about 30% to about 35%, or about 35% to about 40%),an albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, and a thickness of about 5-7 nm (such as about 6 nm) isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the thickness of the albumin coatingof the nanoparticles under cryo-TEM, wherein a percentage of albuminmonomers among the albumin on the nanoparticles being about 40% to about60% (such as about 40% to about 55%, about 40% to about 54%, about 40%to about 53%, about 40% to about 52%, about 40% to about 50%, about 40%to about 48%, or about 40% to about 46%), an albumin to paclitaxel ratioof about 1:2 to about 1:6 in the nanoparticles, and a thickness of about5-7 nm (such as about 6 nm) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles and determining the thickness of the albumin coatingof the nanoparticles under cryo-TEM, wherein a percentage of albuminpolymer among the albumin on the nanoparticles being about 15% to about40% (such as about 15% to about 20%, about 20% to about 24.5%, about24.5% to about 30%, about 30% to about 35%, or about 35% to about 40%),and a percentage of albumin monomer among the albumin on thenanoparticles being at least about 40% to about 60% (such as about 40%to about 55%, about 40% to about 54%, about 40% to about 53%, about 40%to about 52%, about 40% to about 50%, about 40% to about 48%, or about40% to about 46%), an albumin to paclitaxel ratio of about 1:2 to about1:6 in the nanoparticles, and a thickness of about 5-7 nm (such as about6 nm) is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin monomer amongthe albumin on the nanoparticles being less than about 51% is indicativeof suitability of the pharmaceutical composition for medical use. Insome embodiments, a percentage of albumin polymers and oligomers amongthe albumin on the nanoparticles being more than about 35% is indicativeof suitability of the pharmaceutical composition for medical use. Insome embodiments, a percentage of albumin polymers among the albumin onthe nanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe weight ratio of albumin to paclitaxel in the nanoparticles, anddetermining the thickness of the albumin coating of the nanoparticlesunder cryo-TEM, wherein a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, and athickness of about 5-7 nm (such as about 6 nm) is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the method further comprises determining the solubility ofthe pharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the thickness of the albumin coatingof the nanoparticles under cryo-TEM, wherein a percentage of albuminpolymer among the albumin on the nanoparticles being about 15% to about40% (such as about 15% to about 20%, about 20% to about 24.5%, about24.5% to about 30%, about 30% to about 35%, or about 35% to about 40%),a weight percentage of the albumin in the nanoparticles being about 15%to about 30% (such as about 20% to about 25%, about 15% to about 24%, orabout 15% to about 20%), an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles, and a thickness of about 5-7 nm (such asabout 6 nm) is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin monomer among the albumin on the nanoparticles being less thanabout 51% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersand oligomers among the albumin on the nanoparticles being more thanabout 35% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersamong the albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the thickness of the albumin coatingof the nanoparticles under cryo-TEM, wherein a percentage of albuminmonomers among the albumin on the nanoparticles being about 40% to about60% (such as about 40% to about 55%, about 40% to about 54%, about 40%to about 53%, about 40% to about 52%, about 40% to about 50%, about 40%to about 48%, or about 40% to about 46%), a weight percentage of thealbumin in the nanoparticles being about 15% to about 30% (such as about20% to about 25%, about 15% to about 24%, or about 15% to about 20%), analbumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, and a thickness of about 5-7 nm (such as about 6 nm) isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the thickness of the albumin coatingof the nanoparticles under cryo-TEM, wherein a percentage of albuminpolymer among the albumin on the nanoparticles being about 15% to about40% (such as about 15% to about 20%, about 20% to about 24.5%, about24.5% to about 30%, about 30% to about 35%, or about 35% to about 40%),a percentage of albumin monomer among the albumin on the nanoparticlesbeing at least about 40% to about 60% (such as about 40% to about 55%,about 40% to about 54%, about 40% to about 53%, about 40% to about 52%,about 40% to about 50%, about 40% to about 48%, or about 40% to about46%), and a weight percentage of the albumin in the nanoparticles beingabout 15% to about 30% (such as about 20% to about 25%, about 15% toabout 24%, or about 15% to about 20%), an albumin to paclitaxel ratio ofabout 1:2 to about 1:6 in the nanoparticles, and a thickness of about5-7 nm (such as about 6 nm) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe morphology and thickness of the albumin coating of the nanoparticlesunder cryo-TEM, wherein an irregular shape of the nanoparticles and athickness of about 5-7 nm (such as about 6 nm) is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the method further comprises determining the solubility ofthe pharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles and determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), and an irregular shape of the nanoparticles and a thickness ofabout 5-7 nm (such as about 6 nm) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles and determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, wherein a percentage ofalbumin monomers among the albumin on the nanoparticles being about 40%to about 60% (such as about 40% to about 55%, about 40% to about 54%,about 40% to about 53%, about 40% to about 52%, about 40% to about 50%,about 40% to about 48%, or about 40% to about 46%), an irregular shapeof the nanoparticles, and a thickness of about 5-7 nm (such as about 6nm) is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, the method further comprisesdetermining the solubility of the pharmaceutical composition (includingdetermining solubility after storage). In some embodiments, a percentageof albumin monomer among the albumin on the nanoparticles being lessthan about 51% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin polymers and oligomers among the albumin on the nanoparticlesbeing more than about 35% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thepaclitaxel crystallinity of the pharmaceutical composition (for exampleby X-ray diffraction and/or polarized light microscopy, includingdetermining crystallinity after storage). In some embodiments, themethod further comprises determining the paclitaxel recovery following a0.2 micron filtration (including determining recovery after storage). Insome embodiments, the method further comprises determining bindingaffinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles and determining the morphology of the nanoparticles andthickness of the nanoparticles under cryo-TEM, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), a percentage of albumin monomer among the albumin on thenanoparticles being at least about 40% to about 60% (such as about 40%to about 55%, about 40% to about 54%, about 40% to about 53%, about 40%to about 52%, about 40% to about 50%, about 40% to about 48%, or about40% to about 46%), an irregular shape of the nanoparticles, and athickness of about 5-7 nm (such as about 6 nm) is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, anddetermining the morphology of the nanoparticles and thickness of thenanoparticles under cryo-TEM, wherein a weight percentage of the albuminin the nanoparticles being about 15% to about 30% (such as about 20% toabout 25%, about 15% to about 24%, or about 15% to about 20%), anirregular shape of the nanoparticles, and a thickness of about 5-7 nm(such as about 6 nm) is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), a weight percentage of the albumin in the nanoparticles beingabout 15% to about 30% (such as about 20% to about 25%, about 15% toabout 24%, or about 15% to about 20%), an irregular shape of thenanoparticles, and a thickness of about 5-7 nm (such as about 6 nm) isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, wherein a percentage ofalbumin monomers among the albumin on the nanoparticles being about 40%to about 60% (such as about 40% to about 55%, about 40% to about 54%,about 40% to about 53%, about 40% to about 52%, about 40% to about 50%,about 40% to about 48%, or about 40% to about 46%), a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%), an irregular shape of the nanoparticles, and a thickness of about5-7 nm (such as about 6 nm) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), a percentage of albumin monomer among the albumin on thenanoparticles being at least about 40% to about 60% (such as about 40%to about 55%, about 40% to about 54%, about 40% to about 53%, about 40%to about 52%, about 40% to about 50%, about 40% to about 48%, or about40% to about 46%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an irregularshape of the nanoparticles, and a thickness of about 5-7 nm (such asabout 6 nm) is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin monomer among the albumin on the nanoparticles being less thanabout 51% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersand oligomers among the albumin on the nanoparticles being more thanabout 35% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersamong the albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight ratio of albumin to paclitaxel in the nanoparticles,determining the morphology of the nanoparticles and thickness of thealbumin coating under cryo-TEM, wherein an albumin to paclitaxel ratioof about 1:2 to about 1:6 in the nanoparticles, an irregular shape ofthe nanoparticles, and a thickness of about 5-7 nm (such as about 6 nm)is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, the method further comprisesdetermining the solubility of the pharmaceutical composition (includingdetermining solubility after storage). In some embodiments, the methodfurther comprises determining the paclitaxel crystallinity of thepharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesand thickness of the albumin coating under cryo-TEM, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles, an irregular shape of the nanoparticles,and a thickness of about 5-7 nm (such as about 6 nm) is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesand thickness of the albumin coating under cryo-TEM, wherein apercentage of albumin monomers among the albumin on the nanoparticlesbeing about 40% to about 60% (such as about 40% to about 55%, about 40%to about 54%, about 40% to about 53%, about 40% to about 52%, about 40%to about 50%, about 40% to about 48%, or about 40% to about 46%), analbumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, an irregular shape of the nanoparticles, and a thicknessof about 5-7 nm (such as about 6 nm) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesand thickness of the albumin coating under cryo-TEM, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), a percentage of albumin monomer among thealbumin on the nanoparticles being at least about 40% to about 60% (suchas about 40% to about 55%, about 40% to about 54%, about 40% to about53%, about 40% to about 52%, about 40% to about 50%, about 40% to about48%, or about 40% to about 46%), an albumin to paclitaxel ratio of about1:2 to about 1:6 in the nanoparticles, an irregular shape of thenanoparticles, and a thickness of about 5-7 nm (such as about 6 nm) isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe weight ratio of albumin to paclitaxel in the nanoparticles, anddetermining the morphology of the nanoparticles and thickness of thealbumin coating under cryo-TEM, wherein a weight percentage of thealbumin in the nanoparticles being about 15% to about 30% (such as about20% to about 25%, about 15% to about 24%, or about 15% to about 20%), analbumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, an irregular shape of the nanoparticles, and a thicknessof about 5-7 nm (such as about 6 nm) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesand thickness of the albumin coating under cryo-TEM, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, anirregular shape of the nanoparticles, and a thickness of about 5-7 nm(such as about 6 nm) is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, a percentage ofalbumin monomer among the albumin on the nanoparticles being less thanabout 51% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersand oligomers among the albumin on the nanoparticles being more thanabout 35% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, a percentage of albumin polymersamong the albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage). In some embodiments,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesand thickness of the albumin coating under cryo-TEM, wherein apercentage of albumin monomers among the albumin on the nanoparticlesbeing about 40% to about 60% (such as about 40% to about 55%, about 40%to about 54%, about 40% to about 53%, about 40% to about 52%, about 40%to about 50%, about 40% to about 48%, or about 40% to about 46%), aweight percentage of the albumin in the nanoparticles being about 15% toabout 30% (such as about 20% to about 25%, about 15% to about 24%, orabout 15% to about 20%), an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles, an irregular shape of the nanoparticles,and a thickness of about 5-7 nm (such as about 6 nm) is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the morphology of the nanoparticlesand thickness of the albumin coating under cryo-TEM, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), a percentage of albumin monomer among thealbumin on the nanoparticles being at least about 40% to about 60% (suchas about 40% to about 55%, about 40% to about 54%, about 40% to about53%, about 40% to about 52%, about 40% to about 50%, about 40% to about48%, or about 40% to about 46%), a weight percentage of the albumin inthe nanoparticles being about 15% to about 30% (such as about 20% toabout 25%, about 15% to about 24%, or about 15% to about 20%), analbumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, an irregular shape of the nanoparticles, and a thicknessof about 5-7 nm (such as about 6 nm) is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe distribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue; wherein anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700, 800, 900, 1000, 1100 or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, the method further comprises determiningthe solubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the size ofthe nanoparticles (for example by dynamic light scattering). In someembodiments, the method further comprises determining the polydispersityindex of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the span of sizedistribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of the nanoparticles in thepharmaceutical composition. In some embodiments, the method furthercomprises determining the surface potential of the nanoparticles. Insome embodiments, the method further comprises determining thepercentage of the paclitaxel in the nanoparticles among the totalpaclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%) and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the size ofthe nanoparticles (for example by dynamic light scattering). In someembodiments, the method further comprises determining the polydispersityindex of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the span of sizedistribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of the nanoparticles in thepharmaceutical composition. In some embodiments, the method furthercomprises determining the surface potential of the nanoparticles. Insome embodiments, the method further comprises determining thepercentage of the paclitaxel in the nanoparticles among the totalpaclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%) and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, anddetermining the distribution of paclitaxel in a tumor tissue uponinjection of the pharmaceutical composition directly into the tumortissue, wherein a weight percentage of the albumin in the nanoparticlesbeing about 15% to about 30% (such as about 20% to about 25%, about 15%to about 24%, or about 15% to about 20%) and an enhanced paclitaxeltumor distribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, the method further comprises determiningthe solubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%), and an enhanced paclitaxel tumor distribution is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radially for a distance that is greater than (for example morethan about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×,3×, 4×, 5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%) and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thepaclitaxel crystallinity of the pharmaceutical composition (for exampleby X-ray diffraction and/or polarized light microscopy, includingdetermining crystallinity after storage). In some embodiments, themethod further comprises determining the paclitaxel recovery following a0.2 micron filtration (including determining recovery after storage). Insome embodiments, the method further comprises determining bindingaffinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), a weightpercentage of the albumin in the nanoparticles being about 15% to about30% (such as about 20% to about 25%, about 15% to about 24%, or about15% to about 20%), and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight ratio of albumin to paclitaxel in the nanoparticles anddetermining the distribution of paclitaxel in a tumor tissue uponinjection of the pharmaceutical composition directly into the tumortissue, wherein an albumin to paclitaxel ratio of about 1:2 to about 1:6in the nanoparticles and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, the method further comprises determining the solubility ofthe pharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor tissue, wherein a percentage of albumin monomers amongthe albumin on the nanoparticles being about 40% to about 60% (such asabout 40% to about 55%, about 40% to about 54%, about 40% to about 53%,about 40% to about 52%, about 40% to about 50%, about 40% to about 48%,or about 40% to about 46%), an albumin to paclitaxel ratio of about 1:2to about 1:6 in the nanoparticles, and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), analbumin to paclitaxel ratio of about 1:2 to about 1:6, and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700, 800, 900, 1000, 1100 or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe weight ratio of albumin to paclitaxel in the nanoparticles, anddetermining the distribution of paclitaxel in a tumor tissue uponinjection of the pharmaceutical composition directly into the tumortissue, wherein a weight percentage of the albumin in the nanoparticlesbeing about 15% to about 30% (such as about 20% to about 25%, about 15%to about 24%, or about 15% to about 20%), an albumin to paclitaxel ratioof about 1:2 to about 1:6 in the nanoparticles, and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%), an albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor tissue, wherein a percentage of albumin monomers amongthe albumin on the nanoparticles being about 40% to about 60% (such asabout 40% to about 55%, about 40% to about 54%, about 40% to about 53%,about 40% to about 52%, about 40% to about 50%, about 40% to about 48%,or about 40% to about 46%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, and determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), a weightpercentage of the albumin in the nanoparticles being about 15% to about30% (such as about 20% to about 25%, about 15% to about 24%, or about15% to about 20%), an albumin to paclitaxel ratio of about 1:2 to about1:6 in the nanoparticles, and an enhanced paclitaxel tumor distributionis indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, the method further comprisesdetermining the solubility of the pharmaceutical composition (includingdetermining solubility after storage). In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining binding affinity of albumin topaclitaxel in the composition (such as a pharmaceutical composition)(for example by equilibrium dialysis, FTIR, NMR, or a combinationthereof). In some embodiments, the method further comprises determiningthe surface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe morphology of the nanoparticles under cryo-TEM and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein anirregular shape of the nanoparticles and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, the method further comprises determiningthe solubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), an irregular shapeof the nanoparticles, and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%), an irregular shape of the nanoparticles, and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), anirregular shape of the nanoparticles, and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe morphology of the nanoparticles under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein aweight percentage of the albumin in the nanoparticles being about 15% toabout 30% (such as about 20% to about 25%, about 15% to about 24%, orabout 15% to about 20%), an irregular shape of the nanoparticles, and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%), an irregular shape of the nanoparticles, and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an irregularshape of the nanoparticles, and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), a weightpercentage of the albumin in the nanoparticles being about 15% to about30% (such as about 20% to about 25%, about 15% to about 24%, or about15% to about 20%), an irregular shape of the nanoparticles, and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight ratio of albumin to paclitaxel in the nanoparticles,determining the morphology of the nanoparticles under cryo-TEM, anddetermining the distribution of paclitaxel in a tumor tissue uponinjection of the pharmaceutical composition directly into the tumortissue, wherein an albumin to paclitaxel ratio of about 1:2 to about 1:6in the nanoparticles, an irregular shape of the nanoparticles, and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, anirregular shape of the nanoparticles, and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%), an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles, an irregular shape of the nanoparticles,and an enhanced paclitaxel tumor distribution is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radially for a distance that is greater than (for example morethan about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×,3×, 4×, 5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), analbumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, an irregular shape of the nanoparticles, and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe weight ratio of albumin to paclitaxel in the nanoparticles,determining the morphology of the nanoparticles under cryo-TEM, anddetermining the distribution of paclitaxel in a tumor tissue uponinjection of the pharmaceutical composition directly into the tumortissue, wherein a weight percentage of the albumin in the nanoparticlesbeing about 15% to about 30% (such as about 20% to about 25%, about 15%to about 24%, or about 15% to about 20%), an albumin to paclitaxel ratioof about 1:2 to about 1:6 in the nanoparticles, an irregular shape ofthe nanoparticles, and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, the method further comprises determining the solubility ofthe pharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%), an albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, irregular shape of the nanoparticles, and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the paclitaxel recovery following a 0.2 micronfiltration (including determining recovery after storage). In someembodiments, the method further comprises determining binding affinityof albumin to paclitaxel in the composition (such as a pharmaceuticalcomposition) (for example by equilibrium dialysis, FTIR, NMR, or acombination thereof). In some embodiments, the method further comprisesdetermining the surface-to-volume ratio of the nanoparticles in thepharmaceutical composition. In some embodiments, the method furthercomprises determining the percentage of albumin monomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin dimers among the albuminon the nanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, anirregular shape of the nanoparticles, and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a percentage of albumin polymer among thealbumin on the nanoparticles being about 15% to about 40% (such as about15% to about 20%, about 20% to about 24.5%, about 24.5% to about 30%,about 30% to about 35%, or about 35% to about 40%), a percentage ofalbumin monomer among the albumin on the nanoparticles being at leastabout 40% to about 60% (such as about 40% to about 55%, about 40% toabout 54%, about 40% to about 53%, about 40% to about 52%, about 40% toabout 50%, about 40% to about 48%, or about 40% to about 46%), a weightpercentage of the albumin in the nanoparticles being about 15% to about30% (such as about 20% to about 25%, about 15% to about 24%, or about15% to about 20%), an albumin to paclitaxel ratio of about 1:2 to about1:6 in the nanoparticles, an irregular shape of the nanoparticles, andan enhanced paclitaxel tumor distribution is indicative of suitabilityof the pharmaceutical composition for medical use. In some embodiments,the composition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe thickness of the albumin coating of the nanoparticles under cryo-TEMand determining the distribution of paclitaxel in a tumor tissue uponinjection of the pharmaceutical composition directly into the tumortissue, wherein a thickness of about 5-7 nm (such as about 6 nm) and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the thickness of the albumin coating of thenanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), a thickness of about 5-7 nm (such as about 6 nm), and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thepaclitaxel crystallinity of the pharmaceutical composition (for exampleby X-ray diffraction and/or polarized light microscopy, includingdetermining crystallinity after storage). In some embodiments, themethod further comprises determining the paclitaxel recovery following a0.2 micron filtration (including determining recovery after storage). Insome embodiments, the method further comprises determining bindingaffinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the thickness of the albumin coating of thenanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin monomers among the albumin on the nanoparticles being about 40%to about 60% (such as about 40% to about 55%, about 40% to about 54%,about 40% to about 53%, about 40% to about 52%, about 40% to about 50%,about 40% to about 48%, or about 40% to about 46%), a thickness of about5-7 nm (such as about 6 nm), and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, the method further comprises determiningthe solubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the paclitaxel crystallinity of thepharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the thickness of the albumin coating of thenanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), a percentage of albumin monomer among the albumin on thenanoparticles being at least about 40% to about 60% (such as about 40%to about 55%, about 40% to about 54%, about 40% to about 53%, about 40%to about 52%, about 40% to about 50%, about 40% to about 48%, or about40% to about 46%), a thickness of about 5-7 nm (such as about 6 nm), andan enhanced paclitaxel tumor distribution is indicative of suitabilityof the pharmaceutical composition for medical use. In some embodiments,the composition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe thickness of the albumin coating of the nanoparticles undercryo-TEM, and determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor tissue, wherein a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), a thickness ofabout 5-7 nm (such as about 6 nm), and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, the method further comprises determiningthe solubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the thickness of the albumin coating of thenanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), a weight percentage of the albumin in the nanoparticles beingabout 15% to about 30% (such as about 20% to about 25%, about 15% toabout 24%, or about 15% to about 20%), a thickness of about 5-7 nm (suchas about 6 nm), and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the thickness of the albumin coating of thenanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin monomers among the albumin on the nanoparticles being about 40%to about 60% (such as about 40% to about 55%, about 40% to about 54%,about 40% to about 53%, about 40% to about 52%, about 40% to about 50%,about 40% to about 48%, or about 40% to about 46%), a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%), a thickness of about 5-7 nm (such as about 6 nm), and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thepaclitaxel crystallinity of the pharmaceutical composition (for exampleby X-ray diffraction and/or polarized light microscopy, includingdetermining crystallinity after storage). In some embodiments, themethod further comprises determining the paclitaxel recovery following a0.2 micron filtration (including determining recovery after storage). Insome embodiments, the method further comprises determining bindingaffinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the thickness of the albumin coating of thenanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), a percentage of albumin monomer among the albumin on thenanoparticles being at least about 40% to about 60% (such as about 40%to about 55%, about 40% to about 54%, about 40% to about 53%, about 40%to about 52%, about 40% to about 50%, about 40% to about 48%, or about40% to about 46%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), a thickness ofabout 5-7 nm (such as about 6 nm), and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight ratio of albumin to paclitaxel in the nanoparticles,determining the thickness of the albumin coating of the nanoparticlesunder cryo-TEM, and determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor tissue, wherein an albumin to paclitaxel ratio of about1:2 to about 1:6 in the nanoparticles, a thickness of about 5-7 nm (suchas about 6 nm), and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, the method further comprises determining the solubility ofthe pharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), an albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, a thickness of about 5-7 nm (such as about 6 nm), and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin monomers among the albumin on the nanoparticles being about 40%to about 60% (such as about 40% to about 55%, about 40% to about 54%,about 40% to about 53%, about 40% to about 52%, about 40% to about 50%,about 40% to about 48%, or about 40% to about 46%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, athickness of about 5-7 nm (such as about 6 nm), and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), and a percentage of albumin monomer among the albumin on thenanoparticles being at least about 40% to about 60% (such as about 40%to about 55%, about 40% to about 54%, about 40% to about 53%, about 40%to about 52%, about 40% to about 50%, about 40% to about 48%, or about40% to about 46%), an albumin to paclitaxel ratio of about 1:2 to about1:6 in the nanoparticles, a thickness of about 5-7 nm (such as about 6nm), and an enhanced paclitaxel tumor distribution is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radially for a distance that is greater than (for example morethan about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×,3×, 4×, 5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments, apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 51% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thepaclitaxel crystallinity of the pharmaceutical composition (for exampleby X-ray diffraction and/or polarized light microscopy, includingdetermining crystallinity after storage). In some embodiments, themethod further comprises determining the paclitaxel recovery following a0.2 micron filtration (including determining recovery after storage). Insome embodiments, the method further comprises determining bindingaffinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe weight ratio of albumin to paclitaxel in the nanoparticles,determining the thickness of the albumin coating of the nanoparticlesunder cryo-TEM, and determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor tissue, wherein a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, athickness of about 5-7 nm (such as about 6 nm), and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), a weight percentage of the albumin in the nanoparticles beingabout 15% to about 30% (such as about 20% to about 25%, about 15% toabout 24%, or about 15% to about 20%), an albumin to paclitaxel ratio ofabout 1:2 to about 1:6 in the nanoparticles, a thickness of about 5-7 nm(such as about 6 nm), and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin monomers among the albumin on the nanoparticles being about 40%to about 60% (such as about 40% to about 55%, about 40% to about 54%,about 40% to about 53%, about 40% to about 52%, about 40% to about 50%,about 40% to about 48%, or about 40% to about 46%), a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%), an albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, a thickness of about 5-7 nm (such as about 6 nm), and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radically for more than about 700 μm (suchas more than about any of 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or1200 μm) within about 24 hours after the composition (such as apharmaceutical composition) is injected into a tumor tissue (for exampleinjected at the paclitaxel amount of about 12 μg (such as at about 4mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, the method further comprises determining the solubility ofthe pharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a percentage ofalbumin polymer among the albumin on the nanoparticles being about 15%to about 40% (such as about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 30% to about 35%, or about 35% to about40%), a percentage of albumin monomer among the albumin on thenanoparticles being at least about 40% to about 60% (such as about 40%to about 55%, about 40% to about 54%, about 40% to about 53%, about 40%to about 52%, about 40% to about 50%, about 40% to about 48%, or about40% to about 46%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, athickness of about 5-7 nm (such as about 6 nm), and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe morphology and thickness of the albumin coating of the nanoparticlesunder cryo-TEM, and determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor tissue, wherein an irregular shape of the nanoparticlesand a thickness of about 5-7 nm (such as about 6 nm), and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), an irregular shape of the nanoparticles, athickness of about 5-7 nm (such as about 6 nm), and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin monomers among the albumin on the nanoparticlesbeing about 40% to about 60% (such as about 40% to about 55%, about 40%to about 54%, about 40% to about 53%, about 40% to about 52%, about 40%to about 50%, about 40% to about 48%, or about 40% to about 46%), anirregular shape of the nanoparticles, a thickness of about 5-7 nm (suchas about 6 nm), and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the morphology of the nanoparticles andthickness of the nanoparticles under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), a percentage of albumin monomer among thealbumin on the nanoparticles being at least about 40% to about 60% (suchas about 40% to about 55%, about 40% to about 54%, about 40% to about53%, about 40% to about 52%, about 40% to about 50%, about 40% to about48%, or about 40% to about 46%), an irregular shape of thenanoparticles, a thickness of about 5-7 nm (such as about 6 nm), and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe morphology of the nanoparticles and thickness of the nanoparticlesunder cryo-TEM, and determining the distribution of paclitaxel in atumor tissue upon injection of the pharmaceutical composition directlyinto the tumor tissue, wherein a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an irregularshape of the nanoparticles, a thickness of about 5-7 nm (such as about 6nm), and an enhanced paclitaxel tumor distribution is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radially for a distance that is greater than (for example morethan about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×,3×, 4×, 5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an irregularshape of the nanoparticles, a thickness of about 5-7 nm (such as about 6nm), and an enhanced paclitaxel tumor distribution is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radially for a distance that is greater than (for example morethan about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×,3×, 4×, 5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin monomers among the albumin on the nanoparticlesbeing about 40% to about 60% (such as about 40% to about 55%, about 40%to about 54%, about 40% to about 53%, about 40% to about 52%, about 40%to about 50%, about 40% to about 48%, or about 40% to about 46%), aweight percentage of the albumin in the nanoparticles being about 15% toabout 30% (such as about 20% to about 25%, about 15% to about 24%, orabout 15% to about 20%), an irregular shape of the nanoparticles, athickness of about 5-7 nm (such as about 6 nm) and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), a percentage of albumin monomer among thealbumin on the nanoparticles being at least about 40% to about 60% (suchas about 40% to about 55%, about 40% to about 54%, about 40% to about53%, about 40% to about 52%, about 40% to about 50%, about 40% to about48%, or about 40% to about 46%), a weight percentage of the albumin inthe nanoparticles being about 15% to about 30% (such as about 20% toabout 25%, about 15% to about 24%, or about 15% to about 20%), anirregular shape of the nanoparticles, a thickness of about 5-7 nm (suchas about 6 nm), and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight ratio of albumin to paclitaxel in the nanoparticles,determining the morphology of the nanoparticles and thickness of thealbumin coating under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, anirregular shape of the nanoparticles, a thickness of about 5-7 nm (suchas about 6 nm), and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, the method further comprises determining the solubility ofthe pharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles, an irregular shape of the nanoparticles,a thickness of about 5-7 nm (such as about 6 nm), and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin monomers amongthe albumin on the nanoparticles. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin monomers among the albumin on the nanoparticlesbeing about 40% to about 60% (such as about 40% to about 55%, about 40%to about 54%, about 40% to about 53%, about 40% to about 52%, about 40%to about 50%, about 40% to about 48%, or about 40% to about 46%), analbumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, an irregular shape of the nanoparticles, a thickness ofabout 5-7 nm (such as about 6 nm), and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, the method further comprises determiningthe solubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), a percentage of albumin monomer among thealbumin on the nanoparticles being at least about 40% to about 60% (suchas about 40% to about 55%, about 40% to about 54%, about 40% to about53%, about 40% to about 52%, about 40% to about 50%, about 40% to about48%, or about 40% to about 46%), an albumin to paclitaxel ratio of about1:2 to about 1:6 in the nanoparticles, an irregular shape of thenanoparticles, a thickness of about 5-7 nm (such as about 6 nm), and anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe weight ratio of albumin to paclitaxel in the nanoparticles,determining the morphology of the nanoparticles and thickness of thealbumin coating under cryo-TEM, and determining the distribution ofpaclitaxel in a tumor tissue upon injection of the pharmaceuticalcomposition directly into the tumor tissue, wherein a weight percentageof the albumin in the nanoparticles being about 15% to about 30% (suchas about 20% to about 25%, about 15% to about 24%, or about 15% to about20%), an albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, an irregular shape of the nanoparticles, a thickness ofabout 5-7 nm (such as about 6 nm), and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, the method further comprises determiningthe solubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), a weight percentage of the albumin in thenanoparticles being about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%), an albumin topaclitaxel ratio of about 1:2 to about 1:6 in the nanoparticles, anirregular shape of the nanoparticles, a thickness of about 5-7 nm (suchas about 6 nm), and an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows paclitaxel to spread radially for a distance that is greater than(for example more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×,1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of asolvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®) under the sameassay conditions. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if upon tumorinjection it allows paclitaxel to spread radically for more than about700 μm (such as more than about any of 700 μm, 800 μm, 900 μm, 1000 μm,1100 μm or 1200 μm) within about 24 hours after the composition (such asa pharmaceutical composition) is injected into a tumor tissue (forexample injected at the paclitaxel amount of about 12 μg (such as atabout 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor). In someembodiments, a percentage of albumin monomer among the albumin on thenanoparticles being less than about 51% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers and oligomers among the albumin on thenanoparticles being more than about 35% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 17% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 54% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 18% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 55%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers being more than about 65% isindicative of suitability of the pharmaceutical composition for medicaluse. In some embodiments, the method further comprises determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage). In some embodiments, the method furthercomprises determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage). In someembodiments, the method further comprises determining the paclitaxelrecovery following a 0.2 micron filtration (including determiningrecovery after storage). In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin monomers among the albumin on the nanoparticlesbeing about 40% to about 60% (such as about 40% to about 55%, about 40%to about 54%, about 40% to about 53%, about 40% to about 52%, about 40%to about 50%, about 40% to about 48%, or about 40% to about 46%), aweight percentage of the albumin in the nanoparticles being about 15% toabout 30% (such as about 20% to about 25%, about 15% to about 24%, orabout 15% to about 20%), an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles, an irregular shape of the nanoparticles,a thickness of about 5-7 nm (such as about 6 nm), and an enhancedpaclitaxel tumor distribution is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if upon tumor injection it allows paclitaxel to spreadradially for a distance that is greater than (for example more thanabout any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×,5×, 6×, 7×, 8×, or more of) that of a solvent-based paclitaxelformulation (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) under the same assay conditions. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if upon tumor injection it allows paclitaxel tospread radically for more than about 700 μm (such as more than about anyof 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm) within about 24hours after the composition (such as a pharmaceutical composition) isinjected into a tumor tissue (for example injected at the paclitaxelamount of about 12 μg (such as at about 4 mg/ml) into a pancreatic MIAPaCa-2 xenograft tumor). In some embodiments, a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 51%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers andoligomers among the albumin on the nanoparticles being more than about35% is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 17% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 54% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, apercentage of albumin polymers among the albumin on the nanoparticlesbeing more than about 18% and a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 55% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers being more than about 65% is indicative of suitability ofthe pharmaceutical composition for medical use. In some embodiments, themethod further comprises determining the solubility of thepharmaceutical composition (including determining solubility afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel crystallinity of the pharmaceutical composition (forexample by X-ray diffraction and/or polarized light microscopy,including determining crystallinity after storage). In some embodiments,the method further comprises determining the paclitaxel recoveryfollowing a 0.2 micron filtration (including determining recovery afterstorage). In some embodiments, the method further comprises determiningbinding affinity of albumin to paclitaxel in the composition (such as apharmaceutical composition) (for example by equilibrium dialysis, FTIR,NMR, or a combination thereof). In some embodiments, the method furthercomprises determining the surface-to-volume ratio of the nanoparticlesin the pharmaceutical composition. In some embodiments, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin oligomers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin polymers among thealbumin on the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of albumin monomers, dimers,oligomers, or polymers among the total albumin in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering). In some embodiments, the method furthercomprises determining the polydispersity index of the nanoparticles inthe pharmaceutical composition. In some embodiments, the method furthercomprises determining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀)of the nanoparticles in the pharmaceutical composition. In someembodiments, the method further comprises determining the surfacepotential of the nanoparticles. In some embodiments, the method furthercomprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC). In some embodiments,the method further comprises determining the percentage of the albuminthat is in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining the in vitro release kinetics of the composition (such as apharmaceutical composition). In some embodiments, the method furthercomprises determining the stability of the pharmaceutical composition(including determining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight percentage of the albumin in thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, and determining thedistribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue, wherein apercentage of albumin polymer among the albumin on the nanoparticlesbeing about 15% to about 40% (such as about 15% to about 20%, about 20%to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%), a percentage of albumin monomer among thealbumin on the nanoparticles being at least about 40% to about 60% (suchas about 40% to about 55%, about 40% to about 54%, about 40% to about53%, about 40% to about 52%, about 40% to about 50%, about 40% to about48%, or about 40% to about 46%), a weight percentage of the albumin inthe nanoparticles being about 15% to about 30% (such as about 20% toabout 25%, about 15% to about 24%, or about 15% to about 20%), analbumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles, an irregular shape of the nanoparticles, a thickness ofabout 5-7 nm (such as about 6 nm), and an enhanced paclitaxel tumordistribution is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if upontumor injection it allows paclitaxel to spread radially for a distancethat is greater than (for example more than about any of 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifupon tumor injection it allows paclitaxel to spread radically for morethan about 700 μm (such as more than about any of 700 μm, 800 μm, 900μm, 1000 μm, 1100 μm or 1200 μm) within about 24 hours after thecomposition (such as a pharmaceutical composition) is injected into atumor tissue (for example injected at the paclitaxel amount of about 12μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2 xenografttumor). In some embodiments, a percentage of albumin monomer among thealbumin on the nanoparticles being less than about 51% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers and oligomers among thealbumin on the nanoparticles being more than about 35% is indicative ofsuitability of the pharmaceutical composition for medical use. In someembodiments, a percentage of albumin polymers among the albumin on thenanoparticles being more than about 17% and a percentage of albuminmonomer among the albumin on the nanoparticles being less than about 54%is indicative of suitability of the pharmaceutical composition formedical use. In some embodiments, a percentage of albumin polymers amongthe albumin on the nanoparticles being more than about 18% and apercentage of albumin monomer among the albumin on the nanoparticlesbeing less than about 55% is indicative of suitability of thepharmaceutical composition for medical use. In some embodiments, a ratioof albumin on the nanoparticles in the forms of polymers and oligomersto the albumin on the nanoparticles in the form of monomers being morethan about 65% is indicative of suitability of the pharmaceuticalcomposition for medical use. In some embodiments, the method furthercomprises determining the solubility of the pharmaceutical composition(including determining solubility after storage). In some embodiments,the method further comprises determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage). In some embodiments, the method further comprises determiningthe paclitaxel recovery following a 0.2 micron filtration (includingdetermining recovery after storage). In some embodiments, the methodfurther comprises determining binding affinity of albumin to paclitaxelin the composition (such as a pharmaceutical composition) (for exampleby equilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining thesurface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe solubility, paclitaxel crystallinity, and paclitaxel recoveryfollowing a 0.2 micron filtration of the pharmaceutical composition,wherein a solubility of about 50 μg/ml to about 80 μg/ml in a 5% humanalbumin solution, a non-crystalline state, and a recovery of at leastabout 80% is indicative of suitability of the pharmaceutical compositionfor medical use. In some embodiments, the solubility, paclitaxelcrystallinity, and/or paclitaxel recovery are determined after storage(for example after storage for at least about 6 hours, 12 hours, 18hours, 24 hours, 36 hours, 48 hours, or 72 hours, such as at roomtemperature, under refrigerated condition, or at 40° C.). In someembodiments, the paclitaxel crystallinity is determined by X-raydiffraction and/or polarized light microscopy. In some embodiments, themethod further comprises determining binding affinity of albumin topaclitaxel in the composition (such as a pharmaceutical composition)(for example by equilibrium dialysis, FTIR, NMR, or a combinationthereof). In some embodiments, the method further comprises determiningthe surface-to-volume ratio of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin dimers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin oligomers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin polymers among the albumin on thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition. Insome embodiments, the method further comprises determining the particlesize of the nanoparticles (for example by dynamic light scattering). Insome embodiments, the method further comprises determining thepolydispersity index of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, the method further comprisesdetermining the span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of thenanoparticles in the pharmaceutical composition. In some embodiments,the method further comprises determining the surface potential of thenanoparticles. In some embodiments, the method further comprisesdetermining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC). In some embodiments, the method further comprisesdetermining the percentage of the albumin that is in thenon-nanoparticle portion among the total albumin in the pharmaceuticalcomposition (for example by size-exclusion chromatography). In someembodiments, the method further comprises determining the in vitrorelease kinetics of the composition (such as a pharmaceuticalcomposition). In some embodiments, the method further comprisesdetermining the stability of the pharmaceutical composition (includingdetermining stability after storage). In some embodiments, the methodfurther comprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

The different determination steps described above may be carried out invarious combinations in a given method.

For example, in some embodiments, there is provided a method ofassessing suitability of a pharmaceutical composition for medical use ina human individual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin monomers and polymersamong the albumin on the nanoparticles, determining the weightpercentage of the albumin in the nanoparticles, determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage), determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage), and determining the paclitaxel recovery following a 0.2 micronfiltration (including determining recovery after storage). In someembodiments, the method further comprises determining binding affinityof albumin to paclitaxel in the composition (such as a pharmaceuticalcomposition) (for example by equilibrium dialysis, FTIR, NMR, or acombination thereof). In some embodiments, the method further comprisesdetermining the size of the nanoparticles (for example by dynamic lightscattering) and/or size distribution of the nanoparticles. In someembodiments, the method further comprises determining the stability ofthe pharmaceutical composition (including determining stability afterstorage). In some embodiments, the method further comprises determiningtumor distribution of paclitaxel upon administration in vivo (forexample by determining tumor distribution of paclitaxel upon injectionof the pharmaceutical composition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers and polymers among the albumin on thenanoparticles, determining the weight ratio of albumin to paclitaxel inthe nanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage), determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage), and determining the paclitaxel recovery following a 0.2 micronfiltration (including determining recovery after storage). In someembodiments, the method further comprises determining binding affinityof albumin to paclitaxel in the composition (such as a pharmaceuticalcomposition) (for example by equilibrium dialysis, FTIR, NMR, or acombination thereof). In some embodiments, the method further comprisesdetermining the particle size of the nanoparticles (for example bydynamic light scattering) and/or size distribution of the nanoparticles.In some embodiments, the method further comprises determining thestability of the pharmaceutical composition (including determiningstability after storage). In some embodiments, the method furthercomprises determining tumor distribution of paclitaxel uponadministration in vivo (for example by determining tumor distribution ofpaclitaxel upon injection of the pharmaceutical composition directlyinto the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, determiningthe morphology of the nanoparticles and thickness of the albumin coatingunder cryo-TEM, determining the solubility of the pharmaceuticalcomposition (including determining solubility after storage),determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage), anddetermining the paclitaxel recovery following a 0.2 micron filtration(including determining recovery after storage). In some embodiments, themethod further comprises determining binding affinity of albumin topaclitaxel in the composition (such as a pharmaceutical composition)(for example by equilibrium dialysis, FTIR, NMR, or a combinationthereof). In some embodiments, the method further comprises determiningthe size of the nanoparticles (for example by dynamic light scattering)and/or size distribution of the nanoparticles. In some embodiments, themethod further comprises determining the stability of the pharmaceuticalcomposition (including determining stability after storage). In someembodiments, the method further comprises determining tumor distributionof paclitaxel upon administration in vivo (for example by determiningtumor distribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers, dimers, oligomers, and polymersamong the albumin on the nanoparticles, determining the weightpercentage of the albumin in the nanoparticles, determining themorphology of the nanoparticles and thickness of the albumin coatingunder cryo-TEM, determining the surface-to-volume ratio of thenanoparticles in the pharmaceutical composition, and determining thepercentage of albumin monomers, dimers, oligomers, or polymers among thetotal albumin in the pharmaceutical composition. In some embodiments,the method further comprises determining binding affinity of albumin topaclitaxel in the composition (such as a pharmaceutical composition)(for example by equilibrium dialysis, FTIR, NMR, or a combinationthereof). In some embodiments, the method further comprises determiningthe particle size of the nanoparticles (for example by dynamic lightscattering) and/or size distribution of the nanoparticles. In someembodiments, the method further comprises determining the stability ofthe pharmaceutical composition (including determining stability afterstorage). In some embodiments, the method further comprises determiningtumor distribution of paclitaxel upon administration in vivo (forexample by determining tumor distribution of paclitaxel upon injectionof the pharmaceutical composition directly into the tumor tissue).

In some embodiments, there is provided a method of a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin monomers, dimers,oligomers, and polymers among the albumin on the nanoparticles,determining the weight percentage of the albumin in the nanoparticles,determining the paclitaxel crystallinity of the pharmaceuticalcomposition (for example by X-ray diffraction and/or polarized lightmicroscopy, including determining crystallinity after storage),determining the paclitaxel recovery following a 0.2 micron filtration(including determining recovery after storage), determining thepercentage of albumin monomers, dimers, oligomers, or polymers among thetotal albumin in the pharmaceutical composition, determining thepercentage of the paclitaxel in the nanoparticles among the totalpaclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC), and determining the percentage of the albumin thatis in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining binding affinity of albumin to paclitaxel in the composition(such as a pharmaceutical composition) (for example by equilibriumdialysis, FTIR, NMR, or a combination thereof). In some embodiments, themethod further comprises determining the particle size of thenanoparticles (for example by dynamic light scattering) and/or sizedistribution of the nanoparticles. In some embodiments, the methodfurther comprises determining the stability of the pharmaceuticalcomposition (including determining stability after storage). In someembodiments, the method further comprises determining tumor distributionof paclitaxel upon administration in vivo (for example by determiningtumor distribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin monomers, dimers,oligomers, and polymers among the albumin on the nanoparticles,determining the weight percentage of the albumin in the nanoparticles,determining the weight ratio of albumin to paclitaxel in thenanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage), determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage), determining the paclitaxel recovery following a 0.2 micronfiltration (including determining recovery after storage), determiningthe percentage of albumin monomers, dimers, oligomers, or polymers amongthe total albumin in the pharmaceutical composition, determining thepercentage of the paclitaxel in the nanoparticles among the totalpaclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC), and determining the percentage of the albumin thatis in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the method further comprisesdetermining binding affinity of albumin to paclitaxel in the composition(such as a pharmaceutical composition) (for example by equilibriumdialysis, FTIR, NMR, or a combination thereof). In some embodiments, themethod further comprises determining the particle size of thenanoparticles (for example by dynamic light scattering) and/or sizedistribution of the nanoparticles. In some embodiments, the methodfurther comprises determining the stability of the pharmaceuticalcomposition (including determining stability after storage). In someembodiments, the method further comprises determining tumor distributionof paclitaxel upon administration in vivo (for example by determiningtumor distribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of a method of assessingsuitability of a pharmaceutical composition for medical use in a humanindividual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the percentage of albumin monomers, dimers,oligomers, and polymers among the albumin on the nanoparticles,determining the weight percentage of the albumin in the nanoparticles,determining the weight ratio of albumin to paclitaxel in thenanoparticles, determining the morphology of the nanoparticles andthickness of the albumin coating under cryo-TEM, determining thesolubility of the pharmaceutical composition (including determiningsolubility after storage), determining the paclitaxel crystallinity ofthe pharmaceutical composition (for example by X-ray diffraction and/orpolarized light microscopy, including determining crystallinity afterstorage), determining the paclitaxel recovery following a 0.2 micronfiltration (including determining recovery after storage), determiningthe percentage of albumin monomers, dimers, oligomers, or polymers amongthe total albumin in the pharmaceutical composition, determining thepercentage of the paclitaxel in the nanoparticles among the totalpaclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC), determining the percentage of the albumin that isin the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography), determining the particle size of the nanoparticles (forexample by dynamic light scattering) and/or size distribution of thenanoparticles, and determining the stability of the pharmaceuticalcomposition (including determining stability after storage). In someembodiments, the method further comprises determining binding affinityof albumin to paclitaxel in the composition (such as a pharmaceuticalcomposition) (for example by equilibrium dialysis, FTIR, NMR, or acombination thereof). In some embodiments, the method further comprisesdetermining tumor distribution of paclitaxel upon administration in vivo(for example by determining tumor distribution of paclitaxel uponinjection of the pharmaceutical composition directly into the tumortissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe solubility, paclitaxel crystallinity, and paclitaxel recoveryfollowing a 0.2 micron filtration of the pharmaceutical composition,determining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition,determining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC), and determining the percentage of the albumin thatis in the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography). In some embodiments, the solubility, paclitaxelcrystallinity, and/or paclitaxel recovery are determined after storage(for example after storage for at least about 6 hours, 12 hours, 18hours, 24 hours, 36 hours, 48 hours, or 72 hours, such as at roomtemperature, under refrigerated condition, or at 40° C.). In someembodiments, the paclitaxel crystallinity is determined by X-raydiffraction and/or polarized light microscopy. In some embodiments, themethod further comprises determining binding affinity of albumin topaclitaxel in the composition (such as a pharmaceutical composition)(for example by equilibrium dialysis, FTIR, NMR, or a combinationthereof). In some embodiments, the method further comprises determiningthe particle size of the nanoparticles (for example by dynamic lightscattering) and/or size distribution of the nanoparticles. In someembodiments, the method further comprises determining the stability ofthe pharmaceutical composition (including determining stability afterstorage). In some embodiments, the method further comprises determiningtumor distribution of paclitaxel upon administration in vivo (forexample by determining tumor distribution of paclitaxel upon injectionof the pharmaceutical composition directly into the tumor tissue).

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe solubility, paclitaxel crystallinity, and paclitaxel recoveryfollowing a 0.2 micron filtration of the pharmaceutical composition,determining the percentage of albumin monomers, dimers, oligomers, orpolymers among the total albumin in the pharmaceutical composition,determining the percentage of the paclitaxel in the nanoparticles amongthe total paclitaxel in the pharmaceutical composition (for example byreversed-phase HPLC), determining the percentage of the albumin that isin the non-nanoparticle portion among the total albumin in thepharmaceutical composition (for example by size-exclusionchromatography), determining the particle size of the nanoparticles (forexample by dynamic light scattering) and/or size distribution of thenanoparticles, and determining the stability of the pharmaceuticalcomposition (including determining stability after storage). In someembodiments, the solubility, paclitaxel crystallinity, and/or paclitaxelrecovery are determined after storage (for example after storage for atleast about any of 6 hours, 12 hours, 18 hours, 24 v, 36 hours, 48hours, or 72 hours, such as at room temperature, under refrigeratedcondition, or at about 40° C.). In some embodiments, the paclitaxelcrystallinity is determined by X-ray diffraction and/or polarized lightmicroscopy. In some embodiments, the method further comprisesdetermining binding affinity of albumin to paclitaxel in the composition(such as a pharmaceutical composition) (for example by equilibriumdialysis, FTIR, NMR, or a combination thereof). In some embodiments, themethod further comprises determining tumor distribution of paclitaxelupon administration in vivo (for example by determining tumordistribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

Determination of Albumin Oligomeric Status on Nanoparticles

The methods of the present application in some embodiments requiredetermination of oligomeric status (e.g., polymers, monomers, dimers,and/or oligomers) of the albumin on the nanoparticles. The oligomericstatus of the albumin on the nanoparticles may impact the particlestability, solubility, dissolution rate, and in vivo distribution.Further, because albumin-paclitaxel binding is greater for crosslinkedalbumin (namely, albumin polymers, oligomers, and dimers) than albuminmonomers, the oligomeric status of the albumin on the nanoparticles mayalso affect in vivo behavior of the albumin-based paclitaxelnanoparticle composition.

In some embodiments, the oligomeric status of the albumin is determinedby size-exclusion chromatography, such as gel permeation chromatographyor HPLC size-exclusion methods, or polyacrylamide gel electrophoresis(such as sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE). In some embodiments, the oligomeric status is determined byisolating the albumin on the nanoparticles in the pharmaceuticalcomposition by, for example, ultracentrifugation or gel filtrationchromatography, and further analyzing the albumin on the nanoparticlesby, for example, size-exclusion chromatography. The different classes ofalbumins can be determined based on differing retention times ofalbumins when subject to a chromatography (such as size-exclusionchromatography, e.g., gel permeation chromatography). In someembodiments, the different classes of albumins can be determined basedon RRT. In some embodiments, the oligomeric status is determined uponreconstitution of the pharmaceutical composition. In some embodiments,the oligomeric status is determined upon storage of the pharmaceuticalcomposition.

In some embodiments, the size-exclusion chromatography method used iscapable of separating monomeric albumin from dimeric albumin, oligomericalbumin, and polymeric albumin. In some embodiments, the size-exclusionchromatography method used is capable of separating dimeric albumin frommonomeric albumin, oligomeric albumin, and polymeric albumin. In someembodiments, the size-exclusion chromatography method used is capable ofseparating oligomeric albumin from monomeric albumin, dimeric albumin,and polymeric albumin. In some embodiments, the size-exclusionchromatography method used is capable of separating polymeric albuminfrom monomeric albumin, dimeric albumin, and polymeric albumin. In someembodiments, the size-exclusion chromatography method used is capable ofseparating all four categories of albumin on the nanoparticles (e.g.,monomeric, dimeric, oligomeric, polymeric).

In some embodiments, when determining the oligomeric status of thealbumin, the separation range for the size-exclusion chromatography isabout 10,000 daltons to about 500,000 daltons. In some embodiments, thesize-exclusion chromatography is run with a TSKgel G3000 SWXL column. Insome embodiments, the size-exclusion chromatography is run with a columnof TOSOH TSKgel G3000 SWXL, 7.8×300 mm, 5 μm or equivalent. In someembodiments, the size-exclusion chromatography is run with a flow rateof about 1 mL/min. In some embodiments, the size-exclusionchromatography is run at ambient temperature. In some embodiments, thesize-exclusion chromatography is run with a column of TOSOH TSKgel G3000SWXL, 7.8×300 mm, 5 μm or equivalent, at a flow rate of about 1 mL/minat room temperature.

The percentage of the albumin on the nanoparticles that is in the formof a monomer, polymer, dimer, and/or oligomer can be determined bycomparing the amount of monomeric, polymeric, dimeric, and/or oligomericalbumin on the nanoparticles with the total amount of the albumin on thenanoparticles.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of monomericalbumin among the albumin on the nanoparticles is about 40% to about60%. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of monomericalbumin among the albumin on the nanoparticles is about any one of 30%,35%, 40%, 45%, 50%, 55%, 60%, 65%, or 70%. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the percentage of monomeric albumin among the albumin onthe nanoparticles is about any one of 20-30%, 30-40%, 40-50%, 50-60%,60-70%, 70-80%, 20-40%, 40-60%, 60-80%, 20-50%, 50-80%, 35-40%, 40-45%,45-50%, 50-55%, 55-60%, 60-65%, 35-45%, 45-55%, 55-65%, 40-55%, or45-60%.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of dimericalbumin among the albumin on the nanoparticles is about 15% to about 30%(such as about 20% to about 25%, about 15% to about 24%, or about 15% toabout 20%). In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thepercentage of dimeric albumin among the albumin on the nanoparticles isabout any one of 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%,21%, 22%, 23%, 24%, 25%, or 30%. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifthe percentage of dimeric albumin among the albumin on the nanoparticlesis about any of 10-12%, 12-14%, 14-15%, 15-16%, 16-17%, 17-18%, 18-19%,19-20%, 20-21%, 21-23%, 23-25%, 10-15%, 15-20%, 20-25%, 15-17%, 17-19%,15-15.5%, 15.5-16%, 16-16.5%, 16.5-17%, 17-17.5%, 17.5-18%, 18-18.5%,18.5-19%, 19-19.5%, 19.5-20%, 15.5-16.5%, 16.5-17.5%, 17.5-18.5%,18.5-19.5%, 15-16.5%, 16-17.5%, 17-18.5%, 18-19.5%, 16.5-19%, or17.5-20%.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of oligomericalbumin among the albumin on the nanoparticles is about 7% to about 15%.In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of oligomericalbumin among the albumin on the nanoparticles is about any one of 5%,6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%,or 25%. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of oligomericalbumin among the albumin on the nanoparticles is about any one of aboutany one of 5-6%, 6-7%, 7-8%, 8-9%, 9-10%, 10-11%, 11-12%, 12-13%,13-14%, 14-15%, 15-16%, 16-17%, 17-20%, 20-25%, 5-7%, 7-9%, 9-11%,11-13%, 13-15%, 7-10%, 10-13%, 7-12%, 12-15%, 10-15%, or 15-20%.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of polymericalbumin among the albumin on the nanoparticles is about 15% to about 40%(such as about 15% to about 20%, about 20% to about 24.5%, about 24.5%to about 30%, about 30% to about 35%, or about 35% to about 40%). Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the percentage of polymeric albumin amongthe albumin on the nanoparticles is about any one of 15%, 16%, 17%, 18%,19%, 20%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, or 40%. Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the percentage of polymeric albumin amongthe albumin on the nanoparticles is any of about 15% to about 16%, about16% to about 17%, about 17% to about 18%, about 18% to about 19%, about19% to about 20%, about 20% to about 21%, about 21% to about 22%, about22% to about 23%, about 23% to about 24%, about 24% to about 25%, about25% to about 26%, about 26% to about 27%, about 27% to about 28%, about28% to about 29%, about 29% to about 30%, about 30% to about 35%, about35% to about 40%, about 15% to about 18%, about 18% to about 20%, about20% to about 23%, about 23% to about 25%, about 25% to about 30%, about30% to about 40%, about 15% to about 20%, about 20% to about 24.5%,about 24.5% to about 30%, about 15% to about 24.5%, about 15% to about18.5%, or about 15% to about 40% (such as about 15% to about 20%, about20% to about 24.5%, about 24.5% to about 30%, about 30% to about 35%, orabout 35% to about 40%). In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thepercentage of polymeric albumin among the albumin on the nanoparticlesis about 23.6-24.7%, or more than about 29.7%. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the percentage of polymeric albumin among the albumin onthe nanoparticles is about 23.6-24.7%. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the percentage of oligomeric albumin among the albumin onthe nanoparticles is about any one of 10-20%, 20-30%, 30-40%, 15-25%,25-35%, 35-40%, 21-22%, 22-23%, 23-24%, 24-25%, 25-26%, 26-28%, 28-30%,20-23%, 23-25%, 25-30%, 23-23.2%, 23.2-23.4%, 23.4-23.6%, 23.6-23.8%,23.8-24%, 24-24.2%, 24.2-24.4%, 24.4-24.6%, 24.6-24.8%, 24.8-25%,23-23.4%, 23.4-23.8%, 23.8-24.2%, 24.2-24.6%, 24.6%-25%, 23-23.5%,23.5-24%, 24-24.5%, 24.5-25%, 23-23.6%, 23.6-24.2%, 24.2-24.7%, or23.6-24.7%.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of polymericalbumin among the albumin on the nanoparticles is more than about 29.7%.In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of polymericalbumin among the albumin on the nanoparticles is about 30% to about32%, about 32% to about 34%, about 34% to about 36%, about 36% to about38%, or about 38% to about 40%.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about 15% to about 40% (suchas about 15% to about 20%, about 20% to about 24.5%, about 24.5% toabout 30%, about 30% to about 35%, or about 35% to about 40%) of thealbumin on the nanoparticles is in the form of polymers, and about 40%to about 60% of the albumin on the nanoparticles is in the form ofmonomers. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if at least about any one of10%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 22%, 23%, 24%, 25%, ofthe albumin on the nanoparticles is in the form of polymers, and aboutany one of 30%, 35%, 40%, 45%, 50%, 55%, 60%, or 65% of the albumin onthe nanoparticles is in the form of monomers. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if about any one of 10-20%, 20-30%, 30-40%, 15-25%, 25-35%,35-40%, 21-22%, 22-23%, 23-24%, 24-25%, 25-26%, 26-28%, 28-30%, 20-23%,23-25%, 25-30%, 23-23.2%, 23.2-23.4%, 23.4-23.6%, 23.6-23.8%, 23.8-24%,24-24.2%, 24.2-24.4%, 24.4-24.6%, 24.6-24.8%, 24.8-25%, 23-23.4%,23.4-23.8%, 23.8-24.2%, 24.2-24.6%, 24.6%-25%, 23-23.5%, 23.5-24%,24-24.5%, 24.5-25%, 23-23.6%, 23.6-24.2%, 24.2-24.7%, or 23.6-24.7% ofthe albumin on the nanoparticles is in the form of polymers, and aboutany one of 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 20-40%,40-60%, 60-80%, 20-50%, 50-80%, 35-40%, 40-45%, 45-50%, 50-55%, 55-60%,60-65%, 35-45%, 45-55%, 55-65%, 40-55%, or 45-60% of the albumin on thenanoparticles is in the form of monomers. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if about 30% to about 32%, about 32% to about 34%, about 34%to about 36%, about 36% to about 38%, or about 38% to about 40% of thealbumin on the nanoparticles is in the form of polymers, and about anyone of 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 20-40%, 40-60%,60-80%, 20-50%, 50-80%, 35-40%, 40-45%, 45-50%, 50-55%, 55-60%, 60-65%,35-45%, 45-55%, 55-65%, 40-55%, or 45-60% of the albumin on thenanoparticles is in the form of monomers.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about 40% to about 60% (suchas about 40% to about 55%, about 40% to about 54%, about 40% to about53%, or about 40% to about 52%, about 40% to about 50%, about 40% toabout 48%, or about 40% to about 46%) of the albumin on thenanoparticles is in the form of monomers. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if about any one of 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,or 70% of the albumin on the nanoparticles is in the form of monomers.In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about any one of 20-30%,30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 20-40%, 40-60%, 60-80%, 20-50%,50-80%, 35-40%, 40-45%, 45-50%, 50-55%, 55-60%, 60-65%, 35-45%, 45-55%,55-65%, 40-55%, or 45-60% of the albumin on the nanoparticles is in theform of monomers. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if less thanabout 51% of the albumin on the nanoparticles are in the form ofmonomers.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about 7% to about 15% of thealbumin on the nanoparticles is in the form of oligomers. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if about any one of 5%, 6%, 7%, 8%, 9%, 10%,11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or 25% of the albuminon the nanoparticles is in the form of oligomers. In some embodiments,the composition (such as a pharmaceutical composition) is suitable formedical use if about any one of 5-6%, 6-7%, 7-8%, 8-9%, 9-10%, 10-11%,11-12%, 12-13%, 13-14%, 14-15%, 15-16%, 16-17%, 17-20%, 20-25%, 5-7%,7-9%, 9-11%, 11-13%, 13-15%, 7-10%, 10-13%, 7-12%, 12-15%, 10-15%, or15-20% of the albumin on the nanoparticles is in the form of oligomers.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about 15% to about 40% (suchas any of about 15% to about 20%, about 20% to about 24.5%, about 24.5%to about 30%, about 30% to about 35%, or about 35% to about 40%) of thealbumin on the nanoparticles is in the form of polymers, and about 40%to about 60% of the albumin on the nanoparticles is in the form ofmonomers. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about any one of 15%, 16%,17%, 18%, 19%, 20%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, or40% of the albumin on the nanoparticles is in the form of polymers, andabout any one of 30%, 35%, 40%, 45%, 50%, 55%, 60%, or 65% of thealbumin on the nanoparticles is in the form of monomers. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if about any one of 10-20%, 20-30%, 30-40%,15-25%, 25-35%, 35-40%, 21-22%, 22-23%, 23-24%, 24-25%, 25-26%, 26-28%,28-30%, 20-23%, 23-25%, 25-30%, 23-23.2%, 23.2-23.4%, 23.4-23.6%,23.6-23.8%, 23.8-24%, 24-24.2%, 24.2-24.4%, 24.4-24.6%, 24.6-24.8%,24.8-25%, 23-23.4%, 23.4-23.8%, 23.8-24.2%, 24.2-24.6%, 24.6%-25%,23-23.5%, 23.5-24%, 24-24.5%, 24.5-25%, 23-23.6%, 23.6-24.2%,24.2-24.7%, or 23.6-24.7% of the albumin on the nanoparticles is in theform of polymers, and about any one of 20-30%, 30-40%, 40-50%, 50-60%,60-70%, 70-80%, 20-40%, 40-60%, 60-80%, 20-50%, 50-80%, 35-40%, 40-45%,45-50%, 50-55%, 55-60%, 60-65%, 35-45%, 45-55%, 55-65%, 40-55%, or45-60% of the albumin on the nanoparticles is in the form of monomers.In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about 30% to about 32%,about 32% to about 34%, about 34% to about 36%, about 36% to about 38%,or about 38% to about 40% of the albumin on the nanoparticles is in theform of polymers, and about any one of 20-30%, 30-40%, 40-50%, 50-60%,60-70%, 70-80%, 20-40%, 40-60%, 60-80%, 20-50%, 50-80%, 35-40%, 40-45%,45-50%, 50-55%, 55-60%, 60-65%, 35-45%, 45-55%, 55-65%, 40-55%, or45-60% of the albumin on the nanoparticles is in the form of monomers.In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if more than about 35% ofalbumin on the nanoparticles are in the forms of polymers and oligomers.In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if less than about 54%monomers. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if more than about 35% ofalbumin on the nanoparticles are in the forms of polymers and oligomers.In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if less than about 54% of thealbumin on the nanoparticles are in the form of monomers, and more thanabout 11% of the albumin in the nanoparticles are in the form ofpolymers. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if less than about 55% of thealbumin on the nanoparticles are in the form of monomers, and more thanabout 18% of the albumin in the nanoparticles are in the form ofpolymers. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the ratio of albumin on thenanoparticles in the forms of polymers and oligomers to the albumin onthe nanoparticles in the form of monomers is more than about 62%.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if less than about 51%including for example less than about any of 51%, 50%, 49%, 48%, 47%,46%, 45%, 44%, 43%, or 42% albumin on the nanoparticles are in the formof monomers. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if more thanabout 30% including for example more than about any of 30%, 31%, 32%,33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% of the albumin on thenanoparticles are in the forms of polymers and oligomers. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if less than about 51% of the albumin on thenanoparticles are in the form of monomers, and more than about 30% (suchas more than about 35%) of the albumin on the nanoparticles are in theform of polymers and oligomers. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifless than about 54% of the albumin on the nanoparticles are in the formof monomers, and more than about 17% of the albumin in the nanoparticlesare in the form of polymers. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if lessthan about 55% of the albumin on the nanoparticles are in the form ofmonomers, and more than about 18% of the albumin in the nanoparticlesare in the form of polymers. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if theratio of albumin on the nanoparticles in the forms of polymers andoligomers to the albumin on the nanoparticles in the form of monomers ismore than about 65% including for example more than about any of 65%,66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, or 76%.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of albumin onthe nanoparticles in the form of monomers minus the percentage ofalbumin on the nanoparticles in the form of dimers is less than about30%. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the percentage of albumin onthe nanoparticles in the form of monomers minus the percentage ofalbumin on the nanoparticles in the form of dimers is less than aboutany of 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%,43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50%. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the percentage of albumin on the nanoparticles in theform of monomers plus the percentage of albumin on the nanoparticles inthe form of oligomers is less than about 56% to about 58%. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if the percentage of albumin on thenanoparticles in the form of monomers plus the percentage of albumin onthe nanoparticles in the form of oligomers is less than about any of56%, 57, 58, 59%, 60%. 61%. 62%. 63%, 64%, or 65%. In some embodiments,the composition (such as a pharmaceutical composition) is suitable formedical use if the percentage of albumin on the nanoparticles in theform of monomers minus the percentage of albumin on the nanoparticles inthe form of polymers is less than about 20% to about 22%. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if the percentage of albumin on thenanoparticles in the form of monomers minus the percentage of albumin onthe nanoparticles in the form of polymers is less than about any of 20%,21%, 22%, 23%, 24%, 25%, 26%, 27% or 28%. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the percentage of albumin on the nanoparticles in theform of dimers plus the percentage of albumin on the nanoparticles inthe form of polymers is greater than about 42%. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the percentage of albumin on the nanoparticles in theform of dimers plus the percentage of albumin on the nanoparticles inthe form of polymers is greater than about any of 42%, 43%, 44%, 45%,46%, 47%, 48%, 49%, or 50%. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if theratio of albumin on the nanoparticles in the forms of polymers andoligomers to the albumin on the nanoparticles in the form of monomersminus dimers is more than about 88%. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the ratio of albumin on the nanoparticles in the forms ofpolymers and oligomers to the albumin on the nanoparticles in the formof monomers minus dimers is more than about any of 74%, 75%, 76%, 77%,78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%,05%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%,117%, 118%, 119%, or 120%.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the weight ratio of albuminon the nanoparticles in the forms of polymers and oligomers to thealbumin on the nanoparticles in the form of monomers is more than about6:10 including for example more than about any of 6:10, 6.2:10, 6.4:10;6.6:10, 6.8:10, 7.0:10, 7.2:10; 7.4:10; 7.6:10; or 7.8:10. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if the weight ratio of albumin dimers toalbumin monomers on the nanoparticles is more than any of 1.6:10,2.0:10, 2.5:10, 3.0:10, or 3.5:10. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifthe weight ratio of albumin oligomers to albumin monomers on thenanoparticles is more than any of 0.5:10; 1:10, 1.2:10, 1.4:10, or1.6:10. In some embodiments, the weight ratio of albumin polymers toalbumin monomers on the nanoparticles is less than 5.8:10, or 5.7:10. Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the weight ratio of albumin polymers toalbumin dimers on the nanoparticles is less than 40:10, 30:10, or 20:10.In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the weight ratio of albuminoligomers to albumin dimers on the nanoparticles is more than any of3.3:10, 3.5:10, 3.6:10, 3.8:10, 4.0:10, 4.2:10, or 4.4:10. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if the weight ratio of albumin polymers toalbumin oligomers on the nanoparticles is less than any of 120:10,100:10, 80:10, 60:10, or 40:10.

Determination of Weight Percentage of Albumin in the Nanoparticles

The methods of the present application in some embodiments requiredetermination of the weight percentage of the albumin in thenanoparticles.

Generally, to determine the weight percentage of the albumin in thenanoparticles, the amount of the albumin on the nanoparticles and thetotal weight of the nanoparticles can be determined. The amount of thealbumin on the nanoparticles can be determined by, for example,chromatography, such as reversed-phase chromatography, size-exclusionchromatography and/or HPLC size-exclusion chromatography methods,spectrophotometric measurements, or mass spectrometric measurements. Insome embodiments, the method comprises separating the nanoparticles fromthe non-nanoparticle portion by ultracentrifugation or gel filtrationchromatography, followed by analyzing the amount of the albumin on thenanoparticles by, for example, size-exclusion chromatography.Spectrophotometric measurements can be used to determine the amount ofthe albumin on the nanoparticles. In some embodiments, the weightpercentage of the albumin in the nanoparticles is determined uponreconstitution of the pharmaceutical composition. In some embodiments,the weight percentage of the albumin in the nanoparticles is determinedupon storage of the pharmaceutical composition.

In some embodiments, the total weight of the nanoparticles is determinedby addition of the amount of the albumin on the nanoparticles and theamount of paclitaxel in the nanoparticle. Amount of the paclitaxel inthe nanoparticles can be determined by, for example, chromatography,such as reversed-phase high performance liquid chromatography (RP-HPLC),spectrophotometric measurements, or mass spectrometric measurements. Insome embodiments, the method comprises determining the amount of thepaclitaxel in the nanoparticles, for example, by reversed-phase HPLC.

In some embodiments, the amount of the albumin on the nanoparticles andthe amount of the paclitaxel in the nanoparticles are used to determinethe total weight of the nanoparticles. The weight percentage of thealbumin in the nanoparticles can be calculated from the amount of thealbumin on the nanoparticles and the total weight of the nanoparticles.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the weight percentage of thealbumin in the nanoparticles is about 15% to about 30% (such as about20% to about 25%, about 15% to about 24%, or about 15% to about 20%). Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the weight percentage of the albumin inthe nanoparticles is about any one of 10%, 12%, 15%, 15.5%, 16%, 16.5%,17%, 17.5%, 18%, 18.5%, 19%, 19.5%, 20%, 22%, 25%, 30%, 35%, or 40%. Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the weight percentage of the albumin inthe nanoparticles is about any one of 10-12%, 12-14%, 14-15%, 15-16%,16-17%, 17-18%, 18-19%, 19-20%, 20-22%, 22-25%, 10-15%, 15-20%, 20-25%,25-30%, 15-17%, 17-19%, 15-15.5%, 15.5-16%, 16-16.5%, 16.5-17%,17-17.5%, 17.5-18%, 18-18.5%, 18.5-19%, 19-19.5%, 19.5-20%, 15.5-16.5%,16.5-17.5%, 17.5-18.5%, 18.5-19.5%, 15-16.5%, 16-17.5%, 17-18.5%,18-19.5%, 16.5-19%, 17.5-20%, 15-17.5%, 17.5-20%, 20-22.5%, or 22.5-24%.

Determination of Weight Ratio of Albumin to Paclitaxel on theNanoparticles

The methods of the present application in some embodiments requiredetermination of the weight ratio of the albumin on the nanoparticles tothe paclitaxel in the nanoparticles.

Exemplary means for determining the amount of the albumin on thenanoparticles and the amount of the paclitaxel in the nanoparticles arediscussed above. In some embodiments, the weight ratio of albumin topaclitaxel in the nanoparticles is determined by the amount of thealbumin on the nanoparticles over the amount of the paclitaxel in thenanoparticles. In some embodiment, the weight ratio of albumin topaclitaxel in the nanoparticles is determined upon storage of thepharmaceutical composition.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the weight ratio of albuminto paclitaxel in the nanoparticles is about 1:2 to about 1:6. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if the weight ratio of albumin to paclitaxel inthe nanoparticles is about any of 1:1, 1:2, 1:3, 1:3.5, 1:4, 1:4.5, 1:5,1:5.5, 1:6, 1:6.5, 1:7, 1:8, 1:9, or 1:10. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the weight ratio of albumin to paclitaxel in thenanoparticles is any of about 1:1 to about 1:2, about 1:2 to about 1:3,about 1:3 to about 1:3.5, about 1:3.5 to about 1:4, about 1:4 to about1:4.5, about 1:4.5 to about 1:5, about 1:5 to about 1:5.5, about 1:5.5to about 1:6, about 1:6 to about 1:6.5, about 1:6.5 to about 1:7, about1:7 to about 1:8, about 1:8 to about 1:9, about 1:9 to about 1:10, about1:1 to about 1:4, about 1:4 to about 1:6, about 1:6 to about 1:10, about1:3 to about 1:4, about 1:4 to about 1:5, about 1:5 to about 1:6, about1:6 to about 1:7, about 1:3.5 to about 1:4.5, about 1:4.5 to about1:5.5, about 1:5.5 to about 1:6.5, or about 1:2 to about 1:6.

Determination of Paclitaxel Concentration

The methods of present application in some embodiments requiredetermination of the concentration of paclitaxel in the nanoparticleportion of the composition.

The concentration of paclitaxel in the nanoparticle portion of thecomposition (such as a pharmaceutical composition) can be determined bya variety of techniques including an HPLC assay using UV absorbance.Briefly, for example, the nanoparticle portion of composition (such as apharmaceutical composition) is separated from the non-nanoparticleportion of the composition by ultracentrifugation, for example, at50,000 rpm for 41 minutes at 25° C. The supernatant is removed and thepellet is gently washed with water twice. The pellet is then dispersedin a volume of 50:50 acetonitrile:water solution, for example 3.0 ml, bysonication. The sample is further diluted to ensure a homogenoussolution is formed. The sample is analyzed on an HPLC system equippedwith, for example, a Phenomenex, Curosil PFP guard column (4.6 mm×30 mm,5 μm particle size) and a Phenomenex, Curosil PFP analytical column (4.6mm×250 mm, 5 μm particle size), UV absorbance detector, and dataacquisition system. Chromatograms are generated with the UV absorbancedetector set at 228 nm. Comparison to analysis of paclitaxel standardsis used to determine the concentration of paclitaxel in the nanoparticleportion of the composition. In some embodiments, the paclitaxelconcentration in the nanoparticles is determined by the amount ofpaclitaxel in the nanoparticle portion of the composition in the samevolume of the original sample.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the paclitaxel concentrationin the composition (such as a pharmaceutical composition) is about anyof about 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6 mg/ml, 7 mg/ml, 8 mg/ml,9 mg/ml, or 10 mg/ml.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the paclitaxel concentrationin the composition (such as a pharmaceutical composition) is any ofabout 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6 mg/ml, 7 mg/ml, 8 mg/ml, 9mg/ml, or 10 mg/ml. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thepaclitaxel concentration in the composition (such as a pharmaceuticalcomposition) is about 4.4-4.5 mg/ml, 4.5-4.6 mg/ml, 4.6-4.7 mg/ml,4.7-4.8 mg/ml, 4.8-4.9 mg/ml, or 4.9-5.0 mg/ml. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the paclitaxel in the suspension is about 5 mg/ml. Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the paclitaxel in the composition (suchas a pharmaceutical composition) is about 5 mg/ml.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the paclitaxel in thenanoparticle portion of the composition (such as a pharmaceuticalcomposition) is any of about 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, or 10 mg/ml. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the paclitaxel in the nanoparticle portion of thecomposition (such as a pharmaceutical composition) is about 4.2-4.3mg/ml, 4.3-4.4 mg/ml, 4.4-4.5 mg/ml, 4.5-4.6 mg/ml, 4.6-4.7 mg/ml,4.7-4.8 mg/ml, 4.9-5.0 mg/ml.

Determination of Nanoparticle Morphology

The methods of the present application in some embodiments requiredetermination of the nanoparticle morphology. The morphology ofpaclitaxel nanoparticles in an albumin-based paclitaxel nanoparticlecomposition can affect particle solubility, dissolution rate, anddisintegration kinetics.

In some embodiments, the methods comprise determining the shape of thenanoparticles, for example by microscopic methods such as cryo-TEM. Insome embodiments, the methods comprise determining the thickness of thealbumin coating on the nanoparticles, for example by microscopic methodssuch as cryo-TEM. In some embodiments, the methods comprise determiningboth the shape of the nanoparticles and the thickness of the albumincoating on the nanoparticles by microscopic methods, such as cryo-TEM.In some embodiments, the shape of the nanoparticles is determined uponreconstitution of the pharmaceutical composition. In some embodiments,the shape of the nanoparticles is determined upon storage of thepharmaceutical composition.

For example, the composition (such as a pharmaceutical composition) canbe rapidly cooled to cryogenic temperatures following reconstitution ofthe composition (such as a pharmaceutical composition) to form avitreous form of the reconstituted composition which can then beanalyzed. The nanoparticles of the composition (such as a pharmaceuticalcomposition) remain in their native structure during cryo-TEM samplepreparation and image recording. In some embodiments, cryo-TEM recordsthe native structure of the nanoparticles of the composition (such as apharmaceutical composition).

In some embodiments, the thickness of the albumin coating on thenanoparticles is calculated based on measured parameters of thenanoparticles, including for example the albumin-to-paclitaxel ratio ofthe nanoparticles. In some embodiments, the thickness of the albumincoating on the nanoparticles is determined upon reconstitution of thepharmaceutical composition. In some embodiments, the thickness of thealbumin coating on the nanoparticles is determined upon storage of thepharmaceutical composition.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the nanoparticles are ofirregular shape. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thenanoparticles have a non-smooth surface. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the nanoparticles are of irregular shape and have anon-smooth surface. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thenanoparticles have a high degree of rugosity. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use of the nanoparticles are of irregular shape and have a highdegree of rugosity.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the thickness of the albumincoating on the nanoparticles is about 5 nanometers to about 7 nanometersas measured by cryo-TEM. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thenanoparticles are of irregular shape and have an albumin coating with athickness of about 5 nanometers to about 7 nanometers as measured bycryo-TEM. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the albumin coating has athickness of about any of 3 nanometers, 4 nanometers, 5 nanometers, 5.5nanometers, 6 nanometers, 6.5 nanometers, 7 nanometers, 8 nanometers, or9 nanometers as measured by cryo-TEM. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the albumin coating has a thickness of about any of 3-4nanometers, 4-5 nanometers, 5-6 nanometers, 6-7 nanometers, 7-8nanometers, 8-9 nanometers, 3-5 nanometers, 5-7 nanometers, 7-9nanometers, 5-5.5 nanometers, 5.5-6 nanometers, 6-6.5 nanometers, 6.5-7nanometers, 4.5-5.5 nanometers, 5.5-6.5 nanometers, 6.5-7.5 nanometers,5-6.5 nanometers, or 5.5-7 nanometers as measured by cryo-TEM.

In some embodiments, the methods described herein further comprisedetermining the surface-to-volume ratio of the nanoparticles.Surface-to-volume ratios of the nanoparticles can be determined, forexample, by microscopy methods, such as, cryo-TEM, atomic forcemicroscopy, or Fourier transform infrared spectroscopy. In someembodiments, the surface-to-volume ratio of the nanoparticles isdetermined upon reconstitution of the pharmaceutical composition. Insome embodiments, the surface-to-volume ratio of the nanoparticles isdetermined upon storage of the pharmaceutical composition.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the surface-to-volume ratioof the nanoparticles is more than about 46.2:1 μm⁻¹, or thesurface-to-volume ratio of a perfect sphere having the same particlesize as the nanoparticles. In some embodiments, the composition (such asa pharmaceutical composition) is suitable for medical use if thesurface-to-volume ratio of the nanoparticles is more than about any of30:1 μm⁻¹, 35:1 μm⁻¹, 40:1 μm⁻¹, 45:1 μm⁻¹, 46.2:1 μm⁻¹, 47:1 μm⁻¹, 48:1μm⁻¹, 49:1 μm⁻¹, 50:1 μm⁻¹, 52:1 μm⁻¹, 55:1 μm⁻¹, 58:1 μm⁻¹, 60:1 μm⁻¹,65:1 μm⁻¹, 70:1 μm⁻¹, 75:1 μm⁻¹, 80:1 μm⁻¹, 90:1 μm⁻¹, 100:1 μm⁻¹, 110:1μm⁻¹, 120:1 μm⁻¹, 130:1 μm⁻¹, 140:1 μm⁻¹, 150:1 μm⁻¹, 160:1 μm⁻¹, 170:1μm⁻¹, 180:1 μm⁻¹, 190:1 μm⁻¹, 200:1 μm⁻¹, 210:1 μm⁻¹, 220:1 μm⁻¹, 250:1μm⁻¹, 300:1 μm⁻¹, or 400:1 μm⁻¹. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifthe surface-to-volume ratio of the nanoparticles is more than any ofabout 30:1 μm⁻¹ to about 35:1 μm⁻¹, about 35:1 μm⁻¹ to about 40:1 μm⁻¹,about 40:1 μm⁻¹ to about 45:1 μm⁻¹, about 46.2:1 μm⁻¹ to about 47:1μm⁻¹, about 47:1 μm⁻¹ to about 48:1 μm⁻¹, about 48:1 μm⁻¹ to about 49:1μm⁻¹, about 49:1 μm⁻¹ to about 50:1 μm⁻¹, about 50:1 μm⁻¹ to about 52:1μm⁻¹, about 52:1 μm⁻¹ to about 55:1 μm⁻¹, about 55:1 μm⁻¹ to about 58:1μm⁻¹, about 58:1 μm⁻¹ to about 60:1 μm⁻¹, about 60:1 μm⁻¹ to about 65:1μm⁻¹, about 65:1 μm⁻¹ to about 70:1 μm⁻¹, about 70:1 μm⁻¹ to about 75:1μm⁻¹, about 75:1 μm⁻¹ to about 80:1 μm⁻¹, about 46.2:1 μm⁻¹ to about50:1 μm⁻¹, about 50:1 μm⁻¹ to about 60:1 μm⁻¹, about 60:1 μm⁻¹ to about70:1 μm⁻¹, about 70:1 μm⁻¹ to about 80:1 μm⁻¹, about 46.2:1 μm⁻¹ toabout 60:1 μm⁻¹, about 60:1 μm⁻¹ to about 80:1 μm⁻¹, about 50:1 μm⁻¹ toabout 70:1 μm⁻¹, about 48:1 μm⁻¹ to about 52:1 μm⁻¹, about 52:1 μm⁻¹ toabout 65:1 μm⁻¹, about 65:1 μm⁻¹ to about 80:1 μm⁻¹, about 80:1 μm⁻¹ toabout 90:1 μm⁻¹, about 100:1 μm⁻¹ to about 120:1 μm⁻¹, about 120:1 μm⁻¹to about 140:1 μm⁻¹, about 140:1 μm⁻¹ to about 160:1 μm⁻¹, about 160:1μm⁻¹ to about 180:1 μm⁻¹, about 180:1 μm⁻¹ to about 200:1 μm⁻¹, about200:1 μm⁻¹ to about 220:1 μm⁻¹, about 220:1 μm⁻¹ to about 250:1 μm⁻¹,about 250:1 μm⁻¹ to about 300:1 μm⁻¹, about 300:1 μm⁻¹ to about 400:1μm⁻¹, about 30:1 μm⁻¹ to about 40:1 μm⁻¹, about 30:1 μm⁻¹ to about 45:1μm⁻¹, about 80:1 μm⁻¹ to about 120:1 μm⁻¹, about 120:1 μm⁻¹ to about150:1 μm⁻¹, or about 150:1 μm⁻¹ to about 200:1 μm⁻¹.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the surface-to-volume ratioof the nanoparticles is more than about 6/d, wherein d is the averagediameter of the nanoparticles (i.e. the surface-to-volume ratio of aperfect sphere having the same particle size as the nanoparticles). Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the surface-to-volume ratio of thenanoparticles is more than about any of 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6,1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 3, 4, 5, or more than 5 timesof 6/d, wherein d is the average diameter of the nanoparticles.

The surface-to-volume ratio of the nanoparticles is related to theaverage diameter of the nanoparticles. As used herein, “diameter of thenanoparticle” refers to the diameter of the sphere that has the samevolume or weight as the nanoparticle. “Average diameter of thenanoparticles” is the average of the diameters of all nanoparticles inthe composition (such as a pharmaceutical composition). For example, insome embodiments, when the average diameter of the nanoparticles is nomore than about 130 nm, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the surface-to-volume ratioof the nanoparticles is more than about 46.2:1 μm⁻¹. In someembodiments, when the average diameter of the nanoparticles is no morethan about 130 nm, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the surface-to-volume ratioof the nanoparticles is more than about any of 46.2:1 μm⁻¹, 47:1 μm⁻¹,48:1 μm⁻¹, 49:1 μm⁻¹, 50:1 μm⁻¹, 52:1 μm⁻¹, 55:1 μm⁻¹, 58:1 μm⁻¹, 60:1μm⁻¹, 65:1 μm⁻¹, 70:1 μm⁻¹, 75:1 μm⁻¹, 80:1 μm⁻¹, 90:1 μm⁻¹, 100:1 μm⁻¹,110:1 μm⁻¹, 120:1 μm⁻¹, or 140:1 μm⁻¹. In some embodiments, when theaverage diameter of the nanoparticles is no more than about 130 nm, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the surface-to-volume ratio of the nanoparticles is morethan any of about 46.2:1 μm⁻¹ to about 47:1 μm⁻¹, about 47:1 μm⁻¹ toabout 48:1 μm⁻¹, about 48:1 μm⁻¹ to about 49:1 μm⁻¹, about 49:1 μm⁻¹ toabout 50:1 μm⁻¹, about 50:1 μm⁻¹ to about 52:1 μm⁻¹, about 52:1 μm⁻¹ toabout 55:1 μm⁻¹, about 55:1 μm⁻¹ to about 58:1 μm⁻¹, about 58:1 μm⁻¹ toabout 60:1 μm⁻¹, about 60:1 μm⁻¹ to about 65:1 μm⁻¹, about 65:1 μm⁻¹ toabout 70:1 μm⁻¹, about 70:1 μm⁻¹ to about 75:1 μm⁻¹, about 75:1 μm⁻¹ toabout 80:1 μm⁻¹, about 46.2:1 μm⁻¹ to about 50:1 μm⁻¹, about 50:1 μm⁻¹to about 60:1 μm⁻¹, about 60:1 μm⁻¹ to about 70:1 μm⁻¹, about 70:1 μm⁻¹to about 80:1 μm⁻¹, about 80:1 μm⁻¹ to about 90:1 μm⁻¹, about 90:1 μm⁻¹to about 120:1 μm⁻¹, about 120:1 μm⁻¹ to about 140:1 μm⁻¹, about 46.1:1μm⁻¹ to about 60:1 μm⁻¹, about 60:1 μm⁻¹ to about 80:1 μm⁻¹, about 50:1μm⁻¹ to about 70:1 μm⁻¹, about 48:1 μm⁻¹ to about 52:1 μm⁻¹, about 52:1μm⁻¹ to about 65:1 μm⁻¹, or about 65:1 μm⁻¹ to about 80:1 μm⁻¹. In someembodiments, when the average diameter of the nanoparticles is about 60nm to about 190 nm, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the surface-to-volume ratioof the nanoparticles is more than about any of 31.5:1 μm⁻¹, 35:1 μm⁻¹,40:1 μm⁻¹, 45:1 μm⁻¹, 50:1 μm⁻¹, 55:1 μm⁻¹, 60:1 μm⁻¹, 65:1 μm⁻¹, 70:1μm⁻¹, 80:1 μm⁻¹, 90:1 μm⁻¹, 100:1 μm⁻¹, 110:1 μm⁻¹, 120:1 μm⁻¹, 130:1μm⁻¹, 140:1 μm⁻¹, 150:1 μm⁻¹, 160:1 μm⁻¹, 170:1 μm⁻¹, 180:1 μm⁻¹, 190:1μm⁻¹, 200:1 μm⁻¹, or more than 200:1 μm⁻¹. In some embodiments, when theaverage diameter of the nanoparticles is about 120 nm to about 140 nm,the composition (such as a pharmaceutical composition) is suitable formedical use if the surface-to-volume ratio of the nanoparticles is morethan about any of 42.9:1 μm⁻¹, 45:1 μm⁻¹, 50:1 μm⁻¹, 55:1 μm⁻¹, 60:1μm⁻¹, 65:1 μm⁻¹, 70:1 μm⁻¹, 75:1 μm⁻¹, 80:1 μm⁻¹, 85:1 μm⁻¹, 90:1 μm⁻¹,95:1 μm⁻¹, 100:1 μm⁻¹ or more than 100:1 μm⁻¹.

Determination of Particle Size and Polydispersity

The methods of the present application in some embodiments requiredetermination of the size of nanoparticles in the composition (such as apharmaceutical composition). Particle size impacts the dissolution rateof nanoparticles, controls the solubility of nanoparticles, andcontributes to the functional behavior of the nanoparticles.

In some embodiments, the methods comprise determining the size of thenanoparticles in the pharmaceutical composition. In some embodiments,the methods comprise determining the polydispersity index of thenanoparticles in the pharmaceutical composition. In some embodiments,the methods comprise determining the size distribution of thenanoparticles in the pharmaceutical composition. In some embodiments,the polydispersity index of the nanoparticles is determined uponreconstitution of the pharmaceutical composition. In some embodiments,the polydispersity index of the nanoparticles is determined upon storageof the pharmaceutical composition.

In some embodiments, the particle size is determined by laserdiffraction techniques, such as dynamic light scattering. In someembodiments, the size is determined by volume weighted arithmetic meanparticle diameter (D4,3) using a laser diffraction technique. In someembodiments, the particle size is determined by disc centrifugationmethods. In some embodiments, the particle size is determined by tunableresistive pulse sensing (TRPS). In some embodiments, the particle sizeis determined by laser diffraction polarization intensity differentialscattering (PIDS-LD). In some embodiments, the particle size isdetermined by sucrose gradient centrifugation. In some embodiments, theparticle size is determined by analytical centrifugation.

In some embodiments, the polydispersity index is determined by, forexample, dynamic light scattering. Dv₅₀ is the volume-weighted medianparticle diameter.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the average particle size ofthe nanoparticles in the pharmaceutical composition is less than about200 nm. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the average particle size ofthe nanoparticles in the pharmaceutical composition is less than aboutany of 260 nm, 240 nm, 220 nm, 200 nm, 180 nm, 160 nm, 140 nm, 120 nm,100 nm, 80 nm, or 60 nm. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if the averageparticle size of the nanoparticles in the pharmaceutical composition isabout any of 60-80 nm, 80-100 nm, 100-120 nm, 120-140 nm, 140-160 nm,160-180 nm, 180-200 nm, 200-220 nm, 220-240 nm, 240-260 nm, 80-120 nm,120-160 nm, 160-200 nm, 200-240 nm, 50-100 nm, 100-150 nm, 150-200 nm,200-250 nm, 100-105 nm, 105-115 nm, 115-125 nm, 125-135 nm, 135-145 nm,145-155 nm, 155-160 nm, 100-110 nm, 110-120 nm, 120-130 nm, 130-140 nm,140-150 nm, 150-160 nm, 105-125 nm, 125-145 nm, 145-160 nm, 100-130 nm,130-160 nm, 105-135 nm, 135-160 nm, 100-140 nm, 120-160 nm, 110-150 nm,100-150 nm, 105-155 nm, or 100-160 nm. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the average particle size of the nanoparticles in thepharmaceutical composition is about 130 nm.

The parameter ((Dv₉₀−Dv₁₀)/Dv₅₀) describes the span of distribution ofthe particle sizes of the nanoparticles. Dv₅₀ refers to thevolume-weighted median particle diameter. Dv₉₀ refers to the particlediameter where 90% of the volume of all nanoparticles is contained innanoparticles with smaller diameters. Dv₁₀ refers to the particlediameter where 10% of the volume of all nanoparticles is contained innanoparticles with smaller diameters. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the nanoparticles in the pharmaceutical composition havea span of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of about 0.8 to about1.5. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the nanoparticles in thepharmaceutical composition have a span of size distribution((Dv₉₀−Dv₁₀)/Dv₅₀) of about any of 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2,1.3, 1.4, 1.5, 1.6, 1.7, or 1.8. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifthe nanoparticles in the pharmaceutical composition have a sizedistribution ((DV₉₀−DV₁₀/DV₅₀)) of about any of 0.5-0.6, 0.6-0.7,0.7-0.8, 0.8-0.9, 0.9-1, 1-1.1, 1.1-1.2, 1.2-1.3, 1.3-1.4, 1.4-1.5,1.5-1.6, 1.6-1.7, 1.7-1.8, 0.9-1.1, 1.1-1.3, 1.3-1.5, 1.5-1.7, 0.6-0.8,0.8-1, 1-1.2, 1.2-1.4, 1.4-1.6, 1.6-1.8, 0.5-0.8, 0.8-1.1, 1.1-1.4,1.4-1.8, 0.8-1.1, 1.1-1.4, 0.9-1.2, 1.2-1.5, 0.8-1.2, 0.9-1.3, 1-1.4,1.1-1.5, 0.8-1.3, 0.9-1.4, 1-1.5, 0.8-1.4, 0.9-1.5, or 0.8-1.5.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the nanoparticles in thepharmaceutical composition have a polydispersity index of less thanabout 0.3. In some embodiments, the nanoparticles in the pharmaceuticalcomposition have a polydispersity index of less than about any of 0.3,0.25, 0.2, 0.15, 0.1, or 0.05. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifthe nanoparticles in the pharmaceutical composition have apolydispersity index of about any of 0.05-0.07, 0.07-0.09, 0.09-0.11,0.11-0.13, 0.13-0.15, 0.15-0.17, 0.17-0.2, 0.2-0.25, 0.25-0.3,0.05-0.09, 0.09-0.13, 0.13-0.17, 0.17-0.25, 0.06-0.08, 0.08-0.12,0.12-0.16, 0.16-0.18, 0.18-0.22, 0.22-0.28, 0.28-0.3, 0.06-0.12,0.12-0.18, 0.18-0.3, 0.05-0.1, 0.1-0.15, 0.15-0.2, or 0.2-0.3.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) has a size distribution curve similar tothat of nab-paclitaxel sold under the trademark ABRAXANE®.

Determination of Surface Potential

The methods of the present application in some embodiments comprisedetermining the surface potential, such as zeta-potential, of thenanoparticles. Particle surface potential, such as zeta-potential, canplay an important role in preventing the particles from aggregating.

Zeta-potential of the nanoparticles can be determined by techniques,such as, microelectrophoresis, electrophoretic light scattering, ordynamic electrophoretic mobility. In some embodiments, thezeta-potential of the nanoparticles can be determined by tunableresistive pulse sensing (TRPS). In some embodiments, the zeta-potentialof the nanoparticles is determined upon reconstitution of thepharmaceutical composition. In some embodiments, the zeta-potential ofthe nanoparticles is determined upon storage of the pharmaceuticalcomposition.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the nanoparticles in thepharmaceutical composition have a zeta-potential of about −20 mV toabout −35 mV. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thenanoparticles in the pharmaceutical composition have a zeta-potential ofabout −25 mV. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thenanoparticles in the pharmaceutical composition have a zeta-potential ofabout any of −40 mV, −35 mV, −30 mV, −29 mV, −28 mV, −27 mV, −26 mV, −25mV, −24 mV, −23 mV, −22 mV, −21 mV, −20 mV, −15 mV, −10 mV. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if the nanoparticles in the pharmaceuticalcomposition have a zeta-potential of any of about −40 mV to about −35mV, about −35 mV to about −30 mV, about −30 mV to about −25 mV, about−25 mV to about −20 mV, about −20 mV to about −15 mV, about −15 mV toabout −10 mV, about −30 mV to about −28 mV, about −28 mV to about −26mV, about −26 mV to about −24 mV, about −24 mV to about −22 mV, about−22 mV to about −20 mV, about −29 mV to about −27 mV, about −27 mV toabout −25 mV, about −25 mV to about −23 mV, about −23 mV to about −21mV, about −30 mV to about −26 mV, about −26 mV to about −22 mV, about−28 mV to about −24 mV, about −24 mV to about −20 mV, about −30 mV toabout −25 mV, about −25 mV to about −20 mV, or about 30 mV to about −20mV.

Determination of Paclitaxel Crystallinity

The methods of the present application in some embodiments comprisedetermining the crystalline state of paclitaxel in the composition (suchas a pharmaceutical composition). In some embodiments, the methodcomprises determining the crystalline state of paclitaxel by X-raydiffraction. In some embodiments, the method comprises determining thecrystallinity of paclitaxel by light microscopy, such as polarized lightmicroscopy. In some embodiments, the method comprises determining thecrystallinity of paclitaxel by both X-ray diffraction and a polarizedlight microscopy method. In some embodiments, the method comprisesdetermining the crystallinity of paclitaxel by Raman spectroscopy. Insome embodiments, the method comprises determining the crystallinity ofpaclitaxel by second harmonic generation microscopy. In someembodiments, the method comprises determining the crystallinity ofpaclitaxel by X-ray powder diffraction. In some embodiments, the methodcomprises determining the crystallinity of paclitaxel by differentialscanning calorimetry. In some embodiments, the method comprisesdetermining the crystallinity of paclitaxel by thermal gravimetricanalysis. In some embodiments, the method comprises determining thecrystallinity of paclitaxel using one or more technique selected fromthe group consisting of X-ray diffraction, X-ray powder diffraction,light microscopy, polarized light microscopy, Raman spectroscopy, secondharmonic generation microscopy, differential scanning calorimetry, andthermal gravimetric analysis.

In some embodiments, the method comprises determining the crystallinityof paclitaxel by qualitatively determining one or more crystalline formsof paclitaxel. In some embodiments, the method comprises determining thecrystallinity of paclitaxel by qualitatively determining two crystallineforms of paclitaxel. In some embodiments, the method comprisesdetermining the crystallinity of paclitaxel by quantitativelydetermining one or more crystalline forms of paclitaxel. In someembodiments, the method comprises determining the crystallinity ofpaclitaxel by quantitatively determining two crystalline forms ofpaclitaxel. In some embodiments, the method comprises determining thecrystallinity of paclitaxel by qualitatively and quantitativelydetermining one or more crystalline forms of paclitaxel. In someembodiments, the method comprises determining the crystallinity ofpaclitaxel by qualitatively and quantitatively determining twocrystalline forms of paclitaxel.

In some embodiments, the determination of paclitaxel crystallinity isdetermined immediately after reconstitution. In some embodiments, thedetermination of paclitaxel crystallinity is determined after storage,for example after storage for at least about any of 6 hours, 12 hours,18 hours, 24 hours, 36 hours, 40 hours, 44 hours, 48 hours, 52 hours, 56hours, 60 hours, 64 hours, or 72 hours (for example at room temperature,under refrigerated condition, or at about 40° C.).

In some embodiments, nanoparticles are isolated by, for example,ultracentrifugation or gel permeation chromatography. In someembodiments, following ultracentrifugation, the supernatant is decantedand the pellet is washed with water. The isolated nanoparticles are thendried by, for example, lyophilization.

In some embodiments, subsequent analysis of the nanoparticles by X-raydiffraction can determine the physical state of paclitaxel in thenanoparticles. Non-crystalline paclitaxel in the nanoparticles willexhibit broad scattering halos, indicative of an amorphous material(e.g., non-crystalline). Crystalline paclitaxel in the nanoparticleswill exhibit numerous well-defined scattering peaks.

In some embodiments, X-ray powder diffraction of the dried sample isused (alone or in addition to any other method described herein) todetermine the physical state of the paclitaxel in the nanoparticles.Non-crystalline paclitaxel in the nanoparticles will exhibit broadscattering halos, indicative of an amorphous material (e.g.,non-crystalline). Crystalline paclitaxel in the nanoparticles willexhibit numerous well-defined scattering peaks such as sharp scatteringpeaks.

In some embodiments, polarized light microscopy of a reconstitutedsuspension of nanoparticles is used (alone or in addition to any othermethod described herein) to determine the physical state of paclitaxelin the nanoparticles. A birefringence test can be performed with anoptical microscope to determine if the paclitaxel in the nanoparticlesis crystalline or non-crystalline. Absence of birefringence indicatesthat the paclitaxel remained amorphous.

In some embodiments, Raman spectroscopy can be used (alone or inaddition to any other method described herein) to determine the physicalstate of the paclitaxel in the nanoparticles. Crystalline paclitaxel,such as crystalline paclitaxel hydrate, has peaks at 945-947 cm⁻¹, 1320cm⁻¹, 1349 cm⁻¹, 1360 cm⁻¹, 1631-1647 cm⁻¹ and a shoulder on the peak at1715 cm⁻¹.

In some embodiments, differential scanning calorimetry can be used(alone or in addition to any other method described herein) to determinethe physical state of the paclitaxel in the nanoparticles. Thetemperature of thermal transition, e.g., glass transition, can be usedto determine the presence of crystalline paclitaxel in thenanoparticles. For example, amorphous paclitaxel in the nanoparticleshas a thermal transition of 150° C., the crystalline melt of theanhydrous crystal is 213° C., and the solid-state transition in thecrystal hydrate is 166° C.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the paclitaxel in thepharmaceutical composition is non-crystalline. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the paclitaxel in the reconstituted pharmaceuticalcomposition is non-crystalline for at least about any of 6 hours, 12hours, 18 hours, 24 hours, 36 hours, 40 hours, 44 hours, 48 hours, 52hours, 56 hours, 60 hours, 64 hours, or 72 hours. In some embodiments,the composition (such as a pharmaceutical composition) is suitable formedical use if the paclitaxel in the lyophilized pharmaceuticalcomposition is non-crystalline upon storage at about 40° C. for at leastabout any of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months,12 months, 24 months, or more months. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the paclitaxel in the pharmaceutical composition isnon-crystalline upon storage of the composition (such as apharmaceutical composition) at about 4° C. for at least about 24 hours.In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the paclitaxel in thepharmaceutical composition is non-crystalline upon storage of thecomposition (such as a pharmaceutical composition) for at least aboutany of 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 40 hours, 44hours, 48 hours, 52 hours, 56 hours, 60 hours, 64 hours, or 72 hours at,for example at room temperature, under refrigerated condition, or atabout 40° C. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if nocrystalline form of paclitaxel can be detected in the pharmaceuticalcomposition. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if nocrystalline form of paclitaxel can be detected at a limit of 2% of thetotal paclitaxel mass in the pharmaceutical composition. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if less than about 0.01%, 0.008%, 0.005%, or0.003% of the paclitaxel in the pharmaceutical composition is in acrystalline form. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if nocrystalline form of paclitaxel can be detected in the pharmaceuticalcomposition in solution. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if nocrystalline form of paclitaxel can be detected at a limit of 2% of thetotal paclitaxel mass in the pharmaceutical composition in solution. Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if less than 0.01%, 0.008%, 0.005%, or0.003% of the paclitaxel in the pharmaceutical composition in solutionis in a crystalline form.

Determination of Distribution of the Total Albumin and the TotalPaclitaxel Between Nanoparticles and the Non-Nanoparticle Portion

The methods of the present application in some embodiments comprisedetermining the distribution of the total albumin and the totalpaclitaxel between nanoparticles and the non-nanoparticle portion of thecomposition (such as a pharmaceutical composition). In some embodiments,the method comprises determining the distribution of the total albuminbetween the nanoparticles and the non-nanoparticle portion of thecomposition (such as a pharmaceutical composition). In some embodiments,the method comprises determining the distribution of the totalpaclitaxel between the nanoparticles and the non-nanoparticle portion ofthe composition (such as a pharmaceutical composition). In someembodiments, the distribution of the total paclitaxel between thenanoparticles and the non-nanoparticle portion is determined uponreconstitution of the pharmaceutical composition. In some embodiments,the distribution of the total paclitaxel between the nanoparticles andthe non-nanoparticle portion is determined upon storage of thepharmaceutical composition.

The distribution of the total albumin between the nanoparticles and thenon-nanoparticle portion of the composition (such as a pharmaceuticalcomposition) can be determined by measuring the amount of the albumin onthe nanoparticles of the composition (such as a pharmaceuticalcomposition) and/or the amount of the albumin in the non-nanoparticleportion of the composition (such as a pharmaceutical composition).Exemplary methods for measuring the amount albumin in the nanoparticlesand the non-nanoparticle portion are discussed above. In someembodiments, the distribution of the total albumin is determined as apercentage of the total albumin in the composition (such as apharmaceutical composition) in the non-nanoparticle portion of thecomposition (such as a pharmaceutical composition).

In some embodiments, the distribution of the total paclitaxel in thenanoparticles and/or the non-nanoparticle portion of the composition(such as a pharmaceutical composition) can be determined by measuringthe amount of the paclitaxel in the nanoparticles of the composition(such as a pharmaceutical composition) and the amount of the paclitaxelin the non-nanoparticle portion of the composition (such as apharmaceutical composition). Exemplary methods for measuring the amountof the paclitaxel in the nanoparticles and the non-nanoparticle portionare discussed above. In some embodiments, the distribution of the totalpaclitaxel is determined as a percentage of the total paclitaxel in thecomposition (such as a pharmaceutical composition) in the nanoparticleportion of the composition (such as a pharmaceutical composition).

The amount of the total paclitaxel in the composition (such as apharmaceutical composition) associated with nanoparticles can bedetermined by reversed-phase high performance liquid chromatography(RP-HPLC). For example, the nanoparticles can first be isolated byultracentrifugation or gel filtration chromatography. Subsequently, theamount of the paclitaxel in the nanoparticles can then be determined byassaying with quantitative RP-HPLC methods or mass spectrometricmethods. The amount of the paclitaxel measured from the isolatednanoparticles can then be compared with the amount of the totalpaclitaxel in the composition (such as a pharmaceutical composition) tocalculate the percentage of the total paclitaxel in the composition(such as a pharmaceutical composition) that is associated with thenanoparticles. In some embodiments, the amount of the total paclitaxelin the composition (such as a pharmaceutical composition) associatedwith the nanoparticles can be determined by measuring the amount of thepaclitaxel not associated with the nanoparticles. For example, followingultracentrifugation to pellet the nanoparticles, the amount ofpaclitaxel in the resulting supernatant can be assayed by RP-HPLCmethods to determine the amount of paclitaxel in solution (i.e., notassociated with nanoparticles). The amount of the paclitaxel measuredfrom the supernatant and the amount of the total paclitaxel in thecomposition (such as a pharmaceutical composition) can be used tocalculate the percentage of the total paclitaxel in the composition(such as a pharmaceutical composition) that is associated with thenanoparticles.

The quantity of the albumin in the nanoparticles can be determined fromisolated nanoparticles assayed for albumin content by size-exclusionchromatography or HPLC size-exclusion chromatography methods. Thealbumin in the non-nanoparticle portion can be determined by assayingthe supernatant using a similar size-exclusion chromatography method.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if at least about 95% of thetotal paclitaxel in the composition (such as a pharmaceuticalcomposition) are associated with the nanoparticles. In some embodiments,the composition (such as a pharmaceutical composition) is suitable formedical use if at least about any of 99.5%, 99%, 98.5%, 98%, 97.5%. 97%,96.5%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 85%, or 80% of the totalpaclitaxel in the composition (such as a pharmaceutical composition) areassociated with the nanoparticles. In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifabout any of 80-85%, 85-90%, 90-95%, 95-97%, 97-97.5%, 97.5-98%,98-98.5%, 98.5-99%, 99-99.5%, 97-98%, 98-99%, 97.5-98.5%, 98.5-99.5%,97-98.5%, 97-99%, 97-99.5% of the total paclitaxel in the composition(such as a pharmaceutical composition) are associated with thenanoparticles.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if at least about 95% of thetotal albumins in the composition (such as a pharmaceutical composition)are in the non-nanoparticle portion of the composition (such as apharmaceutical composition). In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if at leastabout any of 99.5%, 99%, 98.5%, 98%, 97.5%, 97%, 96.5%, 96%, 95.5%, 95%,94%, 93%, 92%, 91%, 90%, 85%, or 80% of the total albumin in thecomposition (such as a pharmaceutical composition) are in thenon-nanoparticle portion of the composition (such as a pharmaceuticalcomposition). In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if about any of80-85%, 85-90%, 90-95%, 95-96%, 96-96.5%, 96.5-97%, 97-97.5%, 97.5-98%,98-98.5%, 98.5-99%, 99-99.5%, 96-97%, 97-98%, 98-99%, 96.5-97.5%,97.5-98.5%, 98.5-99.5%, 96-98%, 98-99.5%, 96-98.5%, 97-99.5%, or96-99.5% of the total albumin in the composition (such as apharmaceutical composition) are in the non-nanoparticle portion of thecomposition (such as a pharmaceutical composition).

Determination of Albumin Oligomeric Status in the Composition

The methods of the present application in some embodiments comprisedetermining the oligomeric status (e.g., polymers, monomers, dimers,and/or oligomers) of the total albumin in the composition (such as apharmaceutical composition).

The oligomeric forms of the total albumin in the composition (such as apharmaceutical composition) (e.g., monomeric, dimeric, oligomeric,polymeric) can be determined by, for example, size-exclusionchromatography, such as gel permeation chromatography or HPLCsize-exclusion chromatography methods, or polyacrylamide gelelectrophoresis. For example, the oligomeric status can be determined byanalysis of the total albumin in the composition (such as apharmaceutical composition) by size-exclusion chromatography. Thedifferent classes of albumins can be determined based on differingretention time of albumin when subject to a chromatography (such assize-exclusion chromatography). The distribution of the components canbe confirmed by permeation chromatography. In some embodiments, theoligomeric status of the total albumin in the pharmaceutical compositionis determined upon reconstitution of the pharmaceutical composition. Insome embodiments, the oligomeric status of the total albumin in thepharmaceutical composition is determined upon storage of thepharmaceutical composition.

The amount of monomeric albumin in the composition (such as apharmaceutical composition) can be compared with the amount of the totalalbumin in the composition (such as a pharmaceutical composition) tocalculate the percentage of the total albumin in the composition (suchas a pharmaceutical composition) in the form of monomers. The amount ofdimeric albumin in the composition (such as a pharmaceuticalcomposition) can be compared with the amount of the total albumin in thecomposition (such as a pharmaceutical composition) to calculate thepercentage of the total albumin in the composition (such as apharmaceutical composition) in the form of dimers. The amount ofoligomeric albumin in the composition (such as a pharmaceuticalcomposition) can be compared with the amount of the total albumin in thecomposition (such as a pharmaceutical composition) to calculate thepercentage of the total albumin in the composition (such as apharmaceutical composition) in the form of oligomers. The amount ofpolymeric albumin in the composition (such as a pharmaceuticalcomposition) can be compared with the amount of the total albumin in thecomposition (such as a pharmaceutical composition) to calculate thepercentage of the total albumin in the composition (such as apharmaceutical composition) in the form of polymers.

In some embodiments, the separation range for the size-exclusionchromatography is about 10,000 daltons to about 500,000 daltons. In someembodiments, the size-exclusion chromatography is run with a TSKgelG3000 SWXL column. In some embodiments, the size-exclusionchromatography is run with a column of TOSOH TSKgel G3000 SWXL, 7.8×300mm, 5 μm or equivalent. In some embodiments, the size-exclusionchromatography is run with a flow rate of about 1 mL/min. In someembodiments, the size-exclusion chromatography is run at ambienttemperature. In some embodiments, the size-exclusion chromatography isrun with a column of TOSOH TSKgel G3000 SWXL, 7.8×300 mm, 5 μm orequivalent, at a flow rate of about 1 mL/min at room temperature.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if at least about 2% of thetotal albumin in the composition (such as a pharmaceutical composition)is in the form of polymers. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if about 2%to about 5% of the total albumin in the composition (such as apharmaceutical composition) is in the form of polymers. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if at least about any of 1%, 2%, 2.5%, 3%,3.5%, 4%, 4.5%, 5%, 6%, or 8% of the total albumin in the composition(such as a pharmaceutical composition) is in the form of polymers. Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the percentage of polymeric albumin amongthe total albumin in the pharmaceutical composition is about any of1-2%, 2-2.5%, 2.5-3%, 3-3.5%, 3.5-4%, 4-4.5%, 4.5-5%, 5-6%, 6-8%, 2-3%,3-4%, 4-5%, 5-6%, 2.5-3.5%, 3.5-4.5%, 4.5-5.5%, 2-4%, 4-6%, 3-5%,2.5-4.5%, 4.5-6%, 2-3.5%, or 2-5%.

The present application in some embodiments provides albumin-basedpaclitaxel compositions (such as pharmaceutical compositions) suitablefor medical use in which about 75% to about 87% of the total albumin inthe composition (such as a pharmaceutical composition) is in the form ofmonomers. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about any of 70%, 75%, 76%,77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, or 90% of thetotal albumin in the composition (such as a pharmaceutical composition)is in the form of monomers. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if aboutany of 70-75%, 75-76%, 76-77%, 77-78%, 78-79%, 79-80%, 80-81%, 81-82%,82-83%, 83-84%, 84-85%, 85-86%, 86-87%, 87-90%, 75-77%, 77-79%, 79-81%,81-83%, 83-85%, 85-87%, 76-78%, 78-80%, 80-82%, 82-84%, 84-86%, 75-78%,78-81%, 81-84%, 84-87%, 75-80%, 80-87%, 78-84%, 84-87%, 75-85%, 77-87%,or 75-87% of the total albumin in the composition (such as apharmaceutical composition) is in the form of monomers.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about 1% to about 4% of thetotal albumin in the composition (such as a pharmaceutical composition)is in the form of oligomers. In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if aboutany of 0.5%, 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2%, 2.2%, 2.4%, 2.6%, 2.8%, 3%,3.2%, 3.4%, 3.7%, 4%, or 4.5% of the total albumin in the composition(such as a pharmaceutical composition) is in the form of oligomers. Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if about any of 0.5-1%, 1-1.2%, 1.2-1.4%,1.4-1.6%, 1.6-1.8%, 1.8-2%, 2-2.2%, 2.2-2.4%, 2.4-2.6%, 2.6-2.8%,2.8-3%, 3-3.2%, 3.2-3.4%, 3.4-3.7%, 3.7%-4%, 4-4.5%, 1-1.4%, 1.4-1.8%,1.8-2.2%, 2.2-2.6%, 2.6-3%, 3-3.4%, 3.4-4%, 1.2-1.6%, 1.6-2%, 2-2.4%,2.4-2.8%, 2.8-3.2%, 3.2-3.7%, 1-1.8%, 1.8%-2.4%, 2.4-3%, 3-3.7%, 1.4-2%,2-2.6%, 2.6-3.2%, 3.2-4%, 1-2%, 2-3%, 3-4%, 1.5-2.5%, 2.5-3.5%, 3.5-4%,1-2.5%, 2.5-4%, or 1-4% of the total albumin in the composition (such asa pharmaceutical composition) is in the form of oligomers.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about 6% to about 13% of thetotal albumin in the composition (such as a pharmaceutical composition)is in the form of dimers. In some embodiments, the composition (such asa pharmaceutical composition) is suitable for medical use if about anyof 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of thetotal albumin in the composition (such as a pharmaceutical composition)is in the form of dimers. In some embodiments, the composition (such asa pharmaceutical composition) is suitable for medical use if about anyof 3-4%, 4-5%, 5-6%, 6-7%, 7-8%, 8-9%, 9-10%, 10-11%, 11-12%, 12-13%,13-14%, 14-15%, 5.5-6.5%, 6.5-7.5%, 7.5-8.5%, 8.5-9.5%, 9.5-10.5%,10.5-11.5%, 6-8%, 8-10%, 10-12%, 7-9%, 9-11%, 6.5-8.5%, 8.5-10.5%,10.5-13%, 6-9%, 9-13%, 6-10%, 8-13%, or 6-13% of the total albumin inthe composition (such as a pharmaceutical composition) is in the form ofdimers.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if about 2% to about 5% of thetotal albumin in the composition (such as a pharmaceutical composition)is in the form of polymers, and about 75% to about 85% of the totalalbumin in the composition (such as a pharmaceutical composition) in theform of monomers. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if about any of1%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, or 8% of the total albumin inthe composition (such as a pharmaceutical composition) is in the form ofpolymers, and about any of 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, or 90% of the total albumin in the composition(such as a pharmaceutical composition) is in the form of monomers. Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if about any of 1-2%, 2-2.5%, 2.5-3%,3-3.5%, 3.5-4%, 4-4.5%, 4.5-5%, 5-6%, 6-8%, 2-3%, 3-4%, 4-5%, 5-6%,2.5-3.5%, 3.5-4.5%, 4.5-5.5%, 2-4%, 4-6%, 3-5%, 2.5-4.5%, 4.5-6%,2-3.5%, or 2-5% of the total albumin in the composition (such as apharmaceutical composition) is in the form of polymers, and about any of70-75%, 75-76%, 76-77%, 77-78%, 78-79%, 79-80%, 80-81%, 81-82%, 82-83%,83-84%, 84-85%, 85-86%, 86-87%, 87-90%, 75-77%, 77-79%, 79-81%, 81-83%,83-85%, 85-87%, 76-78%, 78-80%, 80-82%, 82-84%, 84-86%, 75-78%, 78-81%,81-84%, 84-87%, 75-80%, 80-87%, 78-84%, 84-87%, 75-85%, 77-87%, or75-87% of the total albumin in the composition (such as a pharmaceuticalcomposition) is in the form of monomers.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the oligomeric status of thetotal albumin in the composition (such as a pharmaceutical composition)do not change significantly upon storage (such as at about 25° C. forabout any of 3 months, 6 months, 9 months, 12 months, 18 months, 24months, or 36 months). In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thepercentage of albumin polymers in the total albumin in the composition(such as a pharmaceutical composition) does not increase by any of 5%,10%, 20%, 30%, 40%, 50%, or more than 50% upon storage (such as at about25° C. for 18 months). In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thepercentage of albumin polymers in the total albumin in the composition(such as a pharmaceutical composition) does not decrease by any of 5%,10%, 20%, 30%, 40%, 50%, or more than 50% upon storage (such as at about25° C. for 18 months). In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thepercentage of albumin monomers in the total albumin in the composition(such as a pharmaceutical composition) does not increase by any of 5%,10%, 15%, 20%, 30%, 40%, 50%, or more than 50% upon storage (such as atabout 25° C. for 18 months). In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if thepercentage of albumin monomers in the total albumin in the composition(such as a pharmaceutical composition) does not decrease by any of 5%,10%, 15%, 20%, 30%, 40%, 50%, or more than 50% upon storage (such as atabout 25° C. for 18 months).

Determination of Recovery of Paclitaxel Following Filtration

The methods of the present application in some embodiments comprisedetermining the recovery of paclitaxel following filtration. Loss ofpaclitaxel following 0.2 micron filtration is a measure of the factionof paclitaxel mass associated with particles large than 200 nm. Thismeasure can be more sensitive to the large particle fraction thanparticle sizing techniques.

In some embodiments, the methods comprise determining the recovery ofpaclitaxel following filtration with a 0.2 micron filter immediatelyafter reconstitution. In some embodiments, the method comprisesdetermining the recovery of paclitaxel following filtration with a 0.2micron filter upon storage (after storage for at least about any of 6hours, 12 hours, 24 hours, 36 hours, 48 hours, 72 hours or more hours(for example under storage at room temperature, under refrigeratedconditions, or under accelerated storage condition (for example at 40°C.)).

The recovery of the paclitaxel in the composition (such as apharmaceutical composition) following 0.2 micron filtration can bedetermined by measuring the amount, such as weight, of paclitaxel in thecomposition (such as a pharmaceutical composition) that passes throughthe 0.2 micron filter. As discussed above, the amount of paclitaxel canbe measured by RP-HPLC techniques. In some embodiments, to determine therecovery, the amount of paclitaxel that remains in the composition (suchas a pharmaceutical composition) following 0.2 micron filtration iscompared with the amount of the total paclitaxel in the composition(such as a pharmaceutical composition) prior to filtration. In someembodiments, the recovery is assessed following storage of thecomposition (such as a pharmaceutical composition) at elevatedtemperatures, for example, about 40° C. for about 24 hours.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) has a paclitaxel recovery of at least aboutany of 80%, 85%, 90%, 95%, 98%, 99%, 99.5% or more following a 0.2micron filtration (for example immediately after reconstitution). Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the composition (such as a pharmaceuticalcomposition) has a paclitaxel recovery of about any of 80-85%, 85-90%,90-95%, 95-98%, 80-90%, 90-98%, 85-95%, 85-98%, 80-95%, 80-98%, 98%-99%,99%-99.5%, or 99.5%-100% following a 0.2 micron filtration (for exampleimmediately after reconstitution).

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) has a paclitaxel recovery of at least aboutany of 80%, 85%, 90%, 95%, 98%, 99%, 99.5%, or more following a 0.2micron filtration after storage for at least about any of 6 hours, 12hours, 24 hours, 36 hours, 48 hours, 72 hours or more hours (for exampleunder storage at room temperature, under refrigerated conditions, orunder accelerated storage condition (for example at about 40° C.)). Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the composition (such as a pharmaceuticalcomposition) has a paclitaxel recovery of about any of 80-85%, 85-90%,90-95%, 95-98%, 80-90%, 90-98%, 85-95%, 85-98%, 80-95%, 80-98%, 98%-99%,99%-99.5%, or 99.5%-100% following a 0.2 micron filtration after storagefor at least about any of 6 hours, 12 hours, 24 hours, 36 hours, 48hours, 72 hours or more hours (for example under storage at roomtemperature, under refrigerated conditions, or under accelerated storagecondition (for example at about 40° C.)).

Determination of Particle Solubility

The methods of the present application in some embodiments comprisedetermining particle solubility of the albumin-based paclitaxelnanoparticle composition. In some embodiments, the solubility isdetermined immediately after reconstitution. In some embodiments, thesolubility is determined upon storage, for example after storage for atleast about any of 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 72hours or more hours (for example under storage at room temperature,under refrigerated conditions, or under accelerated storage condition(for example at 40° C.)).

In some embodiments, the solubility of the pharmaceutical composition isdetermined by performing dynamic light scattering measurements for aseries of concentrations of the composition (such as a pharmaceuticalcomposition). The proportion of intact particles to free paclitaxel is afunction of the solubility of the particles. Thus, as measured by thismethod, the solubility is determined as the concentration below whichparticles are no longer detectable by dynamic light scattering. In someembodiments, the solubility is determined in a 5% human serum albuminsolution. In some embodiments, the solubility is determined in saline.In some embodiments, the solubility is determined in plasma.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) has a solubility of about 50 to about 80μg/ml when reconstituted in a 5% human albumin solution (for exampleimmediately after reconstitution). In some embodiments, the composition(such as a pharmaceutical composition) has a solubility of about any of40 μg/ml, 45 μg/ml, 50 μg/ml, 55 μg/ml, 60 μg/ml, 65 μg/ml, 70 μg/ml, 75μg/ml, 80 μg/ml, 85 μg/ml, 90 μg/ml, or 100 μg/ml when diluted in a 5%human albumin solution (for example immediately after reconstitution).In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) has a solubility of about any of 40-45μg/ml, 45-50 μg/ml, 50-55 μg/ml, 55-60 μg/ml, 60-65 μg/ml, 65-70 μg/ml,70-75 μg/ml, 75-80 μg/ml, 80-85 μg/ml, 85-90 μg/ml, 40-50 μg/ml, 50-60μg/ml, 60-70 μg/ml, 70-80 μg/ml, 80-90 μg/ml, 45-55 μg/ml, 55-65 μg/ml,65-75 μg/ml, 40-55 μg/ml, 55-70 μg/ml, 65-80 μg/ml, 50-70 μg/ml, 60-80μg/ml, 50-80 μg/ml, 65-90 μg/ml, 65-85 μg/ml, 65-95 μg/ml, 65-100 μg/ml,70-85 μg/ml, 70-90 μg/ml, 75-95 μg/ml, or 75-100 μg/ml whenreconstituted in a 5% human albumin solution (for example immediatelyafter reconstitution).

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) has a solubility of about 50 to about 100μg/ml when reconstituted in a 5% human albumin solution after storagefor at least about any of 6 hours, 12 hours, 24 hours, 36 hours, 48hours, 72 hours or more hours (for example under storage at roomtemperature, under refrigerated conditions, or under accelerated storagecondition (for example at about 40° C.)). In some embodiments, thecomposition (such as a pharmaceutical composition) has a solubility ofabout any of 40 μg/ml, 45 μg/ml, 50 μg/ml, 55 μg/ml, 60 μg/ml, 65 μg/ml,70 μg/ml, 75 μg/ml, 80 μg/ml, 85 μg/ml, 90 μg/ml, 100 μg/ml whenreconstituted in a 5% human albumin solution after storage for at leastabout any of 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 72 hoursor more hours (for example under storage at room temperature, underrefrigerated conditions, or under accelerated storage condition (forexample at about 40° C.)). In some embodiments, the composition (such asa pharmaceutical composition) is suitable for medical use if thecomposition (such as a pharmaceutical composition) has a solubility ofabout any of 40-45 μg/ml, 45-50 μg/ml, 50-55 μg/ml, 55-60 μg/ml, 60-65μg/ml, 65-70 μg/ml, 70-75 μg/ml, 75-80 μg/ml, 80-85 μg/ml, 85-90 μg/ml,40-50 μg/ml, 50-60 μg/ml, 60-70 μg/ml, 70-80 μg/ml, 80-90 μg/ml, 45-55μg/ml, 55-65 μg/ml, 65-75 μg/ml, 40-55 μg/ml, 55-70 μg/ml, 65-80 μg/ml,50-70 μg/ml, 60-80 μg/ml, 50-80 μg/ml, 65-90 μg/ml, 65-85 μg/ml, 65-95μg/ml, 65-100 μg/ml, 70-85 μg/ml, 70-90 μg/ml, 75-95 μg/ml, or 75-100μg/ml when reconstituted in a 5% human albumin solution after storagefor at least about any of 6 hours, 12 hours, 24 hours, 36 hours, 48hours, 72 hours or more hours (for example under storage at roomtemperature, under refrigerated conditions, or under accelerated storagecondition (for example at about 40° C.)).

Determination of Albumin Binding Affinity

The methods of the present application in some embodiments comprisedetermining albumin binding affinity to paclitaxel. In some embodiments,the binding affinity is determined by an equilibrium dialysis test. Insome embodiments, the binding affinity is determined by Fouriertransform infrared spectroscopy (FTIR) and/or nuclear magnetic resonance(NMR) analysis. In some embodiments, the binding affinity is determinedby an equilibrium dialysis test and FTIR. In some embodiments, thebinding affinity is determined by an equilibrium dialysis test and NMR.In some embodiments, the binding affinity is determined by anequilibrium dialysis test, FTIR, and NMR. In some embodiments, thealbumin binding affinity to paclitaxel is determined upon reconstitutionof the pharmaceutical composition. In some embodiments, the albuminbinding affinity to paclitaxel is determined upon storage of thepharmaceutical composition.

In some embodiments, the albumin binding affinity to paclitaxel isdetermined by an equilibrium dialysis test. For example, the composition(such as a pharmaceutical composition) is reconstituted with 0.9% salinesolution to create solutions containing 1 mg/mL, 75 μg/mL and 50 μg/mLpaclitaxel. Additionally, a 10% albumin solution is prepared by dilutingAlbumin (Human) USP with 0.9% sodium chloride solution. 200 μL samples,with replicates, are prepared at various paclitaxel concentrations usingan equilibrium dialysis plate with 12 kD molecular weight cutoff insert.In some embodiments, replicate wells of buffer were also included.Subsequently, the plate is sealed and processed on an orbital shaker,heated to 37° C., for 2 hours at 800 rpm. Then, 50 μL is transferredfrom each sample and buffer well to an empty well on a 386-well platefor the paclitaxel assay. Matrix matching was performed by adding 50 μLof 0.9% sodium chloride solution to the wells containing samples on the386-well plate, and by adding 50 μL of albumin solution to the wellscontaining buffer. 100 μL of an internal paclitaxel standard is added toall wells and the plate is then processed on a plate shaker. Contents ofthe plate are then transferred to a filter plate and vacuum filtered.Subsequently, the filtrate was assayed to determine paclitaxelconcentration using reversed-phase liquid chromatography massspectrometry. Briefly, 10 μL of the filtrate was injected from anautosampler onto a reversed-phase C18 column. Elution is performed usinga gradient method at a flow rate of 0.5 mL/min. Mobile phasecompositions are as follows: A, water with 0.1% formic acid and B,acetonitrile with 0.1% formic acid. Effluent is introduced into MSsystem through heated electrospray ionization.

In some embodiments, the albumin binding affinity to paclitaxel isdetermined by an equilibrium dialysis test after dialysis for at least12 hours, 24 hours, 36 hours, 48 hours, 72 hours, or more hours.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the binding affinity of thealbumin in the composition (such as a pharmaceutical composition) forpaclitaxel is about the same as the binding affinity of albumin in thenab-paclitaxel sold under the trademark ABRANXANE®. In some embodiments,the composition (such as a pharmaceutical composition) is suitable formedical use if the equilibrium binding (or dissociation) constant of thealbumin in the composition (such as a pharmaceutical composition) forpaclitaxel is less than about any of 130 μM, 110 μM, 90 μM, 70 μM, 60μM, 55 μM, 50 μM or 45 μM. In some embodiments, the composition (such asa pharmaceutical composition) is suitable for medical use if the bindingaffinity of the albumin in the composition (such as a pharmaceuticalcomposition) for paclitaxel is about 42 μM.

Determination of In Vitro Release Kinetics

The methods of the present application in some embodiments furthercomprise determining the in vitro release kinetics of the albumin-basedpaclitaxel nanoparticle composition. In some embodiments, thedetermination is carried out immediately after reconstitution. In someembodiments, the determination is carried out upon storage, for exampleupon storage for least about any of 6 hours, 12 hours, 24 hours, 36hours, 48 hours, 72 hours, or more hours (for example under storage atroom temperature, under refrigerated conditions, or under acceleratedstorage condition (for example at about 40° C.)). In some embodiments,the determination is carried out upon storage, for example upon storagefor least about any of 1 month, 2 months, 3 months, 4 months, 5 months,6 months, 7 months, 8 months, 9 months, or more months (for exampleunder storage at room temperature, under refrigerated conditions, orunder accelerated storage condition (for example at about 40° C.)). Insome embodiments, the determination is carried out immediately afterdilution of the composition. In some embodiments, the determination iscarried out after, for example, at least 1 hour, 2 hours, 3 hours, 4hours, 5 hours, 6 hours, or more hours after dilution of thecomposition.

In vitro release kinetics may be assayed using different techniques. Inone example, an in vitro release kinetics assay measures the particlesize and intensity of light scattered by the particles using, forexample, dynamic light scattering over a period of time immediatelyfollowing a reduction in particle concentration. In some embodiments,the release kinetics are determined by diluting the composition (such asa pharmaceutical composition) in a 0.9% saline solution. In someembodiments, the release kinetics is determined by diluting thecomposition (such as a pharmaceutical composition) in a 5% human serumalbumin solution. In a second example, an in vitro release kineticsassay measures the absorbance of the composition over a period of timeimmediately following a reduction in particle concentration. In someembodiments, the absorbance of the composition is measured using aUV-Vis spectrophotometer. In some embodiments, the absorbance of thecomposition is measured using a UV-Vis spectrophotometer equipped with a295 nm cut-off filter. In some embodiments, the absorbance of thecomposition is measured at, for example, 340 nm. In some embodiments,the composition is diluted to, for example, 100 μg/ml, as measured bythe concentration of paclitaxel.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the dissolution profile ofthe composition (such as a pharmaceutical composition), as measured byan in vitro release kinetics assay, matches the dissolution profile ofnab-paclitaxel sold under the trademark ABRAXANE®. In some embodiments,the composition (such as a pharmaceutical composition) is suitable formedical use if the dissolution profile of the composition (such as apharmaceutical composition) following an accelerated aging processmatches the dissolution profile of the nab-paclitaxel sold under thetrademark ABRAXANE®. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thedissolution profile of the composition (such as a pharmaceuticalcomposition) following an accelerated aging process matches thedissolution profile of nab-paclitaxel sold under the trademark ABRAXANE®following the same accelerated aging process. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the composition (such as a pharmaceutical composition)has an in vitro release kinetic behavior that is similar to that of thenab-paclitaxel sold under the trademark ABRAXANE® under the same assayconditions upon storage for at least about any of 6 hours, 12 hours, 24hours, 36 hours, 48 hours, 72 hours, or more hours (for example understorage at room temperature, under refrigerated conditions, or underaccelerated storage condition (for example at about 40° C.)).

In some embodiments, the in vitro release is determined using thedynamic light scattering method. In some embodiments, the in vitrorelease kinetics assay measures the intensity of light scattered by thecomposition over a period of time immediately following a reduction inparticle concentration. In some embodiments, the light scatteringintensity of the composition is measured using a dynamic lightscattering apparatus, where the concentration of paclitaxel releasedfrom the nanoparticles is calculated from the intensity of scatteredlight. In some embodiments, the intensity of light scattered is measuredat a scattering angle of 173°. In some embodiments, the composition isdiluted to 20 μg/ml, as measured by the concentration of paclitaxel, andin some embodiments the composition is diluted to 37.5 μg/ml. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if the mean value of percent releasedpaclitaxel from a nanoparticle composition (such as a pharmaceuticalcomposition) at 20 μg/ml in 10 mM sodium chloride, as measured by thepaclitaxel concentration, after 120 minutes is about 100%. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if the mean value of percent releasedpaclitaxel from a nanoparticle composition (such as a pharmaceuticalcomposition) at 37.5 μg/ml in 10 mM sodium chloride, as measured by thepaclitaxel concentration, after 120 minutes is about 85%.

Determination of Physical Stability

The methods of the present application in some embodiments comprisedetermining the physical stability of particles in the composition (suchas a pharmaceutical composition). In some embodiments, the stability isdetermined immediately after reconstitution. In some embodiments, thestability is measured upon storage, for example upon storage for atleast about any of 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 72hours, or more hours (for example under storage at room temperature,under refrigerated conditions, or under accelerated storage condition(for example at about 40° C.)). In some embodiments, the stability ismeasured after storage, for example, for 8 hours at about 5° C. followedby storage for 8 hours at about 25° C., or after for storage for 24hours at about 25° C.

Stability of nanoparticles in the pharmaceutical composition can beassayed by a number of techniques, including, but not limited to, visualinspection (such as visual appearance, visual color, visible particulatematter), microscopy imaging, and loss of potency. In some embodiments,the stability of the composition (such as a pharmaceutical composition)is determined based on sedimentation. Sedimentation can be assessed byvisual inspection and/or microscopy (such as cross-polarizationmicroscopy). Microscopy can be used to determine the size of aggregatedsediment particles. In some embodiments, the stability of thecomposition (such as a pharmaceutical composition) is determined basedon the crystalline state of the nanoparticles, for example by increasedpresence of nanoparticles with crystalline paclitaxel. In someembodiments, the stability of the composition (such as a pharmaceuticalcomposition) is determined based on a loss of potency following a 0.2micron filtration of the composition (such as a pharmaceuticalcomposition). In some embodiments, the loss of potency is in vitropotency. In some embodiments, the loss of potency is in vivo potency.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the nanoparticles arephysically stable (for example immediately after reconstitution). Insome embodiments, the composition (such as a pharmaceutical composition)is suitable for medical use if the nanoparticles are physically stablefor at least about any of 6 hours, 12 hours, 24 hours, 36 hours, 48hours, 72 hours or more hours upon storage (for example under storage atroom temperature, under refrigerated conditions, or under acceleratedstorage condition (for example at about 40° C.)). In some embodiments,the composition (such as a pharmaceutical composition) is suitable formedical use if the nanoparticles are physically stable after storage for8 hours at about 5° C. followed by storage for 8 hours at about 25° C.,or after for storage for 24 hours at about 25° C.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) shows no visible particulate matter (forexample immediately after reconstitution). In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the composition (such as a pharmaceutical composition)shows no visible particulate matter for at least about any of 6 hours,12 hours, 24 hours, 36 hours, 48 hours, 72 hours or more hours uponstorage (for example under storage at room temperature, underrefrigerated conditions, or under accelerated storage condition (forexample at about 40° C.)).

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) shows no visible particulate matter afterstorage for 8 hours at about 5° C. followed by storage for 8 hours atabout 25° C., or after for storage for 24 hours at about 25° C.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) shows no sedimentation (for exampleimmediately after reconstitution). In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifthe composition (such as a pharmaceutical composition) shows nosedimentation for at least about any of 6 hours, 12 hours, 24 hours, 36hours, 48 hours, 72 hours or more hours upon storage (for example understorage at room temperature, under refrigerated conditions, or underaccelerated storage condition (for example at about 40° C.)). In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if the composition (such as a pharmaceuticalcomposition) shows no sedimentation after storage for 8 hours at about5° C. followed by storage for 8 hours at about 25° C., or after forstorage for 24 hours at about 25° C.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the paclitaxel in thecomposition (such as a pharmaceutical composition) shows nocrystallinity (e.g., by polarized light microscopy) (for exampleimmediately after reconstitution). In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifthe paclitaxel in the composition (such as a pharmaceutical composition)shows no crystallinity (e.g., by polarized light microscopy) for atleast about any of 6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 72hours or more hours upon storage (for example under storage at roomtemperature, under refrigerated conditions, or under accelerated storagecondition (for example at about 40° C.)). In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if the paclitaxel in the composition (such as apharmaceutical composition) shows no crystallinity (e.g., by polarizedlight microscopy) after storage for 8 hours at about 5° C. followed bystorage for 8 hours at about 25° C., or after for storage for 24 hoursat about 25° C.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) shows no loss of potency (e.g., by in vitroor in vivo testing) following a 0.2 micron filtration (for exampleimmediately after reconstitution). In some embodiments, the composition(such as a pharmaceutical composition) is suitable for medical use ifthe composition (such as a pharmaceutical composition) shows no loss ofpotency (e.g., by in vitro or in vivo testing) for at least about any of6 hours, 12 hours, 24 hours, 36 hours, 48 hours, 72 hours or more hoursupon storage (for example under storage at room temperature, underrefrigerated conditions, or under accelerated storage condition (forexample at about 40° C.)). In some embodiments, the composition (such asa pharmaceutical composition) is suitable for medical use if thecomposition (such as a pharmaceutical composition) shows no loss ofpotency (e.g., by in vitro or in vivo testing) after storage for 8 hoursat about 5° C. followed by storage for 8 hours at about 25° C., or afterfor storage for 24 hours at about 25° C.

Determination of Osmolality

The methods described herein in some embodiments comprise determiningthe osmolality of a reconstituted composition (such as a pharmaceuticalcomposition). In some embodiments, the osmolality is determinedimmediately after reconstitution. In some embodiments, the osmolality ismeasured upon storage, for example upon storage for at least about anyof 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months, at atemperature of, for example, about 25° C. or about 40° C. In someembodiments, the osmolality is measured after storage for 6 months atabout 25° C. and a relative humidity of about 60% at an invertedposition. In some embodiments, the osmolality is measured after storagefor 6 months at about 40° C. and a relative humidity of about 75% at aninverted position.

Osmolality of a composition (such as a pharmaceutical composition) canbe assayed by a number of methods, including, comprising use of a vaporpressure depression osmometer, a membrane osmometer, or a freezing pointdepression osmometer.

In some embodiments, the composition (such as a pharmaceuticalcomposition) has an osmolality of between about 320 mOsm/kg to about 360mOsm/kg. In some embodiments, the composition (such as a pharmaceuticalcomposition) has an osmolality of between about 325 mOsm/kg to about 355mOsm/kg. In some embodiments, the composition (such as a pharmaceuticalcomposition) has an osmolality of between about 330 mOsm/kg to about 350mOsm/kg. In some embodiments, the composition (such as a pharmaceuticalcomposition) has an osmolality of about 330 mOsm/kg, 331 mOsm/kg, 332mOsm/kg, 333 mOsm/kg, 334 mOsm/kg, 335 mOsm/kg, 336 mOsm/kg, 337mOsm/kg, 338 mOsm/kg, 339 mOsm/kg, 340 mOsm/kg, 341 mOsm/kg, 342mOsm/kg, 343 mOsm/kg, 344 mOsm/kg, 345 mOsm/kg, 346 mOsm/kg, 347 mOsm/kgor 348 mOsm/kg.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) shows no significant change in osmolalityafter storage. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thecomposition (such as a pharmaceutical composition) shows no significantchange in osmolality after storage at elevated temperatures, such as 40°C. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) shows no significant change in osmolalityfor at least about any of 1 month, 2 months, 3 months, 4 months, 5months, 6 months or more months upon storage (for example under storageat room temperature, under refrigerated conditions, or under acceleratedstorage condition (for example at about 40° C.)).

Determination of Viscosity

The methods described herein in some embodiments comprise determiningthe viscosity (such as dynamic viscosity) of a reconstituted composition(such as a pharmaceutical composition). In some embodiments, theviscosity is determined immediately after reconstitution. In someembodiments, the viscosity is measured upon storage, for example uponstorage for at least about any of 1 month, 2 months, 3 months, 4 months,5 months, or 6 months, at a temperature of, for example, about 25° C. orabout 40° C. In some embodiments, the viscosity is measured afterstorage for 6 months at about 25° C. and a relative humidity of about60% at an inverted position. In some embodiments, the viscosity ismeasured after storage for 6 months at about 40° C. and a relativehumidity of about 75% at an inverted position.

Viscosity (such as dynamic viscosity) of a composition (such as apharmaceutical composition) can be assayed by a number of methods,including, comprising use of a viscometers and rheometers.

In some embodiments, the composition (such as a pharmaceuticalcomposition) has a viscosity of between about 1.20 centipoise to about1.50 centipoise. In some embodiments, the composition (such as apharmaceutical composition) has a viscosity of between about 1.25centipoise to about 1.45 centipoise. In some embodiments, thecomposition (such as a pharmaceutical composition) has a viscosity ofabout 1.25 centipoise, 1.26 centipoise, 1.27 centipoise, 1.28centipoise, 1.29 centipoise, 1.30 centipoise, 1.31 centipoise, 1.32centipoise, 1.33 centipoise, 1.34 centipoise, 1.35 centipoise, 1.36centipoise, 1.37 centipoise, 1.38 centipoise, 1.39 centipoise, 1.40centipoise, 1.41 centipoise, 1.42 centipoise, 1.43 centipoise, 1.44centipoise, or 1.45 centipoise.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) shows no significant change in viscosityafter storage. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if thecomposition (such as a pharmaceutical composition) shows no significantchange in viscosity after storage at elevated temperatures, such as 40°C. In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if the composition (such as apharmaceutical composition) shows no significant change in viscosity forat least about any of 1 month, 2 months, 3 months, 4 months, 5 months, 6months or more months upon storage (for example under storage at roomtemperature, under refrigerated conditions, or under accelerated storagecondition (for example at about 40° C.)).

Determination of Tumor Distribution of Paclitaxel In Vivo

The methods described herein in some embodiments comprise determiningtumor distribution of paclitaxel upon administration of thealbumin-based paclitaxel nanoparticle composition in vivo. Variations inalbumin-based paclitaxel nanoparticle compositions, and theirformulations, can affect in vivo distribution of paclitaxel within tumortissue. In some embodiments, tumor penetration of a composition (such asa pharmaceutical composition) is determined. In some embodiments, tumorcell uptake of a composition (such as a pharmaceutical composition) isdetermined. In some embodiments, tumor penetration and tumor cell updateof a composition (such as a pharmaceutical composition) are determined.

In some embodiments, the method comprises administering the composition(such as a pharmaceutical composition) to an animal having a tumor andmeasuring the level of paclitaxel in the tumor tissue. The level ofpaclitaxel can be measured, for example, by imaging,immunohistochemistry or quantitative mass spectrometry methods. In someembodiments, the paclitaxel in the composition (such as a pharmaceuticalcomposition) that is administered into the animal is radio-labeled, thusallowing sensitive determination of tumor distribution of thepaclitaxel.

In some embodiments, methods for determining the distribution ofpaclitaxel within tumor tissue comprise use of a tumor model. In someembodiments, the methods are performed on a naturally occurring tumor inan individual. In some embodiments, the methods are performed on axenograft tumor in an individual. In some embodiments, the tumor modelis a MIA PaCa-2 xenograft, A2058 xenograft, or H2122 xenograft. In someembodiments, the distribution of paclitaxel within tumor tissue isdetermined upon reconstitution of the pharmaceutical composition. Insome embodiments, the distribution of paclitaxel within tumor tissue isdetermined upon storage of the pharmaceutical composition.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is administered to the animal by systemic administration.In some embodiments, the composition (such as a pharmaceuticalcomposition) is delivered to the tumor by direct injection. In someembodiments, the composition (such as a pharmaceutical composition) isdelivered to the tumor by direct microinjection, for example, via aCIVO™ arrayed microinjection device (Presage Biosciences, Seattle,Wash.). In some embodiments, the composition (such as a pharmaceuticalcomposition) is delivered into flank tumor xenografts. For methodscomprising the use of direct injection, in some embodiments, an imagingagent is injected with the composition (such as a pharmaceuticalcomposition) to locate the injection site. In some embodiments, thecomposition (such as a pharmaceutical composition) is co-injected withan imaging agent, such as a fluorescent compound, for example,VivoTag680-S. In some embodiments, a labeled-paclitaxel is used in thecomposition (such as a pharmaceutical composition).

In some embodiments, methods for determining the distribution ofpaclitaxel within tumor tissue comprise measuring spatial distributionof paclitaxel within the tumor and/or tumor-associated tissue. In someembodiments, spatial distribution of paclitaxel is measured fromsections (e.g., slices) of tumor tissue. In some embodiments, thesections of tumor tissue are perpendicular to the injection path. Insome embodiments, spatial distribution of paclitaxel is measured byimaging, such as immunohistochemical imaging, fluorescent imaging, orany combinations thereof, of the tumor tissue. In some embodiments,spatial distribution of paclitaxel is measured by immunohistochemicalstaining of cells in mitotic arrest. In some embodiments, cells inmitotic arrest are detected by phospho-histone H3 (pHH3) staining, suchas immunohistochemical staining of pHH3 using an anti-pHH3 antibody.Generally, a cell stained by a pHH3 detection agent is indicative of acell in mitotic arrest. In some embodiments, tumor tissue is stainedwith a cell imaging agent, such as an agent for imaging a cell nucleus,for example, 4′,6-diamindino-2-phenylindole (DAPI). In some embodiments,spatial distribution of paclitaxel comprises use of software, such asCIVOanalyzer™ (Presage Biosciences, Seattle, Wash.). In someembodiments, spatial distribution of paclitaxel is measured byperforming mass spectrometry-based imaging (e.g., MALDI-MS-basedimaging). In some embodiments, spatial distribution of paclitaxel ismeasured by detecting labeled-paclitaxel.

In some embodiments, spatial distribution of paclitaxel in a tumortissue is performed on more than one tumor sections (e.g., slices) froma tumor. In some embodiments, spatial distribution of paclitaxel intumor tissue is performed on more than one adjacent tumor sections froma tumor, thereby allowing for the recreation of a 3-dimensional tumordistribution of paclitaxel. In some embodiments, the more than oneadjacent tumor sections are in spatial proximity to a microinjectionsite.

In some embodiments, distribution of paclitaxel within tumor tissue isdetermined by measuring spatial distribution of paclitaxel over atime-course following administration of the composition (such as apharmaceutical composition). In some embodiments, distribution ofpaclitaxel within tumor tissue is measured at about 24 hours, about 48hours, and/or about 72 hours following administration. In someembodiments, distribution of paclitaxel within tumor tissue is measuredat least about 12 hours, at least about 24 hours, at least about 48hours, and/or at least about 72 hours following administration. In someembodiments, distribution of paclitaxel within tumor tissue isdetermined by direct microinjection of a pharmaceutical composition intoflank human tumor xenografts and analyzed, for example, at about 24hours, about 48 hours, and/or about 72 hours following directmicroinjection.

In some embodiments, the distribution of paclitaxel within tumor tissueis determined based on detection of paclitaxel in an area of the tumortissue. In some embodiments, the distribution of paclitaxel within tumortissue is determined based on detection of paclitaxel within a cell.Generally, methods comprising identification of cells in mitotic arrestcomprise assessing the number of cells in mitotic arrest and/or thenumber of cells not in mitotic arrest. In some embodiments, thedistribution of paclitaxel is determined based on the percentage ofcells in mitotic arrest. In some embodiments, the percentage of mitoticarrest is reported as the fraction of pHH3 positive cells (i.e., numberof pHH3 positive cells over the number of total cells in the samplearea). In some embodiments, mitotic arrest is measured at one or moredefined radial distances extending from the site of injection, forexample, at about 100 μm, about 200 μm, about 300 μm, about 400 μm,about 500 μm, about 600 μm, about 700 μm, about 800 μm, about 900 μm,about 1000 μm, about 1100 μm, about 1200 μm, and/or about 1300 μm fromthe site of the injection. In some embodiments, mitotic arrest ismeasured at one or more defined radial distances extending from the siteof injection, for example, at least about 100 μm, at least about 200 μm,at least about 300 μm, at least about 400 μm, at least about 500 μm, atleast about 600 μm, at least about 700 μm, at least about 800 μm, atleast about 900 μm, at least about 1000 μm, at least about 1100 μm, atleast about 1200 μm, and/or at least about 1300 μm from the site of theinjection.

In some embodiments, the distribution of paclitaxel within tumor tissuecomprises determining an area of response (e.g., an area of tumor tissuewherein paclitaxel is detected).

In some embodiments, the distribution profile of paclitaxel within tumortissue for a composition (such as a pharmaceutical composition) iscompared to the distribution profile of paclitaxel for anothercomposition. Generally, the amount (e.g., concentration) of paclitaxelin each composition is measured, for example by mass spectrometry, toallow for normalization of paclitaxel concentrations betweencompositions (such as pharmaceutical compositions).

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if it allows an enhancedpaclitaxel tumor distribution. A composition allows “enhanced paclitaxeltumor distribution” if, upon tumor injection, it allows a distributionof paclitaxel within the tumor tissue that is more extensive than thatof a solvent-based paclitaxel formulation (such as the solvent-basedpaclitaxel formulation sold under the trademark TAXOL®). In someembodiments, the radial distance of paclitaxel spread (penetrationand/or tumor cell uptake of paclitaxel) in a tumor tissue extending fromthe site of injection of the composition (such as a pharmaceuticalcomposition) is greater than (for example more than about 1.1×, 1.2×,1.3×, 1.4×, 1.5×, 1.6×, 1.7×, 1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or moreof) that of a solvent-based paclitaxel formulation (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®)under the same assay conditions.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if upon tumor injection itallows a distribution of paclitaxel within the tumor tissue that issimilar to that of nab-paclitaxel sold under the trademark ABRAXANE®under the same assay conditions. For example, in some embodiments, theradial distance of paclitaxel penetration (or tumor cell uptake ofpaclitaxel) in a tumor tissue extending from the site of injection ofthe composition (such as a pharmaceutical composition) is greater thanor equal to that of nab-paclitaxel sold under the trademark ABRAXANE®under the same assay conditions. In some embodiments, the radialdistance of paclitaxel penetration (or tumor cell uptake of paclitaxel)in a tumor tissue extending from the site of injection of thecomposition (such as a pharmaceutical composition) is at least about 90%(including for example at least about any of 90%, 95%, 98% or 99%) ofthat of nab-paclitaxel sold under the trademark ABRAXANE® under the sameassay conditions.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if it allows paclitaxel tospread radially for more than about 700 μm (such as more than about 1200μm) within about 24 hours when the composition (such as a pharmaceuticalcomposition) is injected into a tumor tissue (for example when injectedat the paclitaxel amount of about 12 μg (or 4 mg/ml) into a pancreaticMIA PaCa-2 xenograft tumor). In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if itallows paclitaxel to spread radially for more than about 700 μm withinabout any one of 12 hours, 24 hours, 36 hours, 48 hours, or 72 hourswhen the composition (such as a pharmaceutical composition) is injectedinto a tumor tissue. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if it allowspaclitaxel to spread radially for more than about 1200 μm within aboutany one of 12 hours, 24 hours, 36 hours, 48 hours, or 72 hours when thecomposition (such as a pharmaceutical composition) is injected into atumor tissue. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if it allowspaclitaxel to spread radially for more than about any of 200 μm, 300 μm,400 μm, 500 μm, 600 μm, 700 μm, 750 μm, 800 μm, 850 μm, 900 μm, 950 μm,1000 μm, 1100 μm, 1200 μm, 1300 μm, 1400 μm, 1500 μm or 2000 μm withinabout 12 hours when the composition (such as a pharmaceuticalcomposition) is injected into a tumor tissue. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if it allows paclitaxel to spread radially for more thanabout any of 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 750 μm, 800μm, 850 μm, 900 μm, 950 μm, 1000 μm, 1100 μm, 1200 μm, 1300 μm, 1400 μm,1500 μm or 2000 μm within about 24 hours when the composition (such as apharmaceutical composition) is injected into a tumor tissue. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if it allows paclitaxel to spread radially formore than about any of 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm,750 μm, 800 μm, 850 μm, 900 μm, 950 μm, 1000 μm, 1100 μm, 1200 μm, 1300μm, 1400 μm, 1500 μm or 2000 μm within about 36 hours when thecomposition (such as a pharmaceutical composition) is injected into atumor tissue. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if it allowspaclitaxel to spread radially for more than about any of 200 μm, 300 μm,400 μm, 500 μm, 600 μm, 700 μm, 750 μm, 800 μm, 850 μm, 900 μm, 950 μm,1000 μm, 1100 μm, 1200 μm, 1300 μm, 1400 μm, 1500 μm or 2000 μm withinabout 48 hours when the composition (such as a pharmaceuticalcomposition) is injected into a tumor tissue. In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if it allows paclitaxel to spread radially for more thanabout any of 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 750 μm, 800μm, 850 μm, 900 μm, 950 μm, 1000 μm, 1100 μm, 1200 μm, 1300 μm, 1400 μm,1500 μm or 2000 μm within about 72 hours when the composition (such as apharmaceutical composition) is injected into a tumor tissue. In someembodiments, the composition (such as a pharmaceutical composition)meets two or more of the above-described functional attributes.

Determination of the radial distance of paclitaxel penetration (or tumorcell uptake) extending from the injection site in a tumor tissuerequires setting a baseline percentage (or fraction) of mitoticallyarrested cells among all cells in the tumor tissue. The radial distanceof paclitaxel penetration (or tumor cell uptake) extending from theinjection site is the radial distance extending from the injection site,beyond which the percentage of mitotically arrested cells among allcells at the radial distance is no more than the baseline percentage. Insome embodiments, the baseline percentage is set to be about 0.6%. Insome embodiments, the baseline percentage is set to be about any of0.1%, 0.3%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%,4%, 5%, 10%, or more. The radial distance of paclitaxel penetration (ortumor cell uptake) extending from the injection site of the composition(such as a pharmaceutical composition), and that of nab-paclitaxel soldunder the trademark ABRAXANE® are compared to determine whether tumordistribution of the composition (such as a pharmaceutical composition)in vivo is similar to that of nab-paclitaxel sold under the trademarkABRAXANE®. Similarly, the radial distance of paclitaxel penetration (ortumor cell uptake) extending from the injection site of the composition(such as a pharmaceutical composition), and that of a solvent-basedpaclitaxel (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) are compared to determine whether tumordistribution of the composition (such as a pharmaceutical composition)in vivo is greater than that of a solvent-based paclitaxel (such as thesolvent-based paclitaxel formulation sold under the trademark TAXOL®).

In some embodiments, tumor distribution of paclitaxel is determinedbased on the percentage of mitotically arrested cells (such as pHH3+cells) at a defined radial distance extending from the site ofinjection. The percentages (or fractions) of mitotically arrested cells(such as pHH3+ cells) at defined radial distances extending from thesite of injection of the compositions (such as pharmaceuticalcompositions) vary according to the volume and dose of the paclitaxel inthe composition (such as a pharmaceutical composition) being injected,the time at which the tumor tissue is harvested, the cell type of thetumor tissue or xenograft (such as cell lines or source of tumortissue), the animal model, or other assay conditions.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if, upon injection of thepharmaceutical composition, the percentage (or fraction) of mitoticallyarrested cells (such as pHH3+ cells) at a radial distance extending fromthe site of injection (such as at any of 200 μm, 400 μm, 500 μm, 600 μm,700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm, 1200 μm, 1500 μm, or 2000 μm)of the composition (such as a pharmaceutical composition) is similar tothat of nab-paclitaxel sold under the trademark ABRAXANE® within about24 hours of injection. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if, uponinjection of the pharmaceutical composition, the percentages (orfractions) of mitotically arrested cells (such as pHH3+ cells) at aplurality of radial distances extending from the site of injection (suchas at any combination of 200 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm,900 μm, 1000 μm, 1100 μm, 1200 μm, 1500 μm, or 2000 μm) of thecomposition (such as a pharmaceutical composition) is similar to thoseof nab-paclitaxel sold under the trademark ABRAXANE® within about 24hours of injection. In some embodiments, the composition (such as apharmaceutical composition) is suitable for medical use if, uponinjection of the pharmaceutical composition, the percentage (orfraction) of mitotically arrested cells (such as pHH3+ cells) at aradial distance extending from the site of injection (such as at any of200 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm, 900 μm, 1000 μm, 1100μm, 1200 μm, 1500 μm, or 2000 μm) of the composition (such as apharmaceutical composition) is greater than that of a solvent-basedpaclitaxel (such as the solvent-based paclitaxel formulation sold underthe trademark TAXOL®) within about 24 hours of injection. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for medical use if, upon injection of the pharmaceuticalcomposition, the percentages (or fractions) of mitotically arrestedcells (such as pHH3+ cells) at a plurality of radial distances extendingfrom the site of injection (such as at any combination of 200 μm, 400μm, 500 μm, 600 μm, 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm, 1200 μm,1500 μm, or 2000 μm) of the composition (such as a pharmaceuticalcomposition) are greater than those of a solvent-based paclitaxel (suchas the solvent-based paclitaxel formulation sold under the trademarkTAXOL®) within about 24 hours of injection.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if it allows more than about 5%(such as more than about any of 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, or 7% ormore) of all cells at a radial distance of about 400 μm extending from asite of injection to be mitotically arrested cells within about 24 hoursof the injection when the composition (such as a pharmaceuticalcomposition) is injected into a tumor tissue (for example when injectedat the paclitaxel amount of about 12 μg (or 4 mg/ml) into a pancreaticMIA PaCa-2 xenograft tumor). In some embodiments, the composition (suchas a pharmaceutical composition) is suitable for medical use if itallows more than about 3.5% (such as more than about any one of 2%,2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5% or more) of all cells at a radialdistance of about 400 μm extending from a site of injection to bemitotically arrested cells within about 24 hours of the injection whenthe composition (such as a pharmaceutical composition) is injected intoa tumor tissue (for example when injected at the paclitaxel amount ofabout 12 μg (or 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor).In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if it allows more than about2.5% (such as more than about any one of 1%, 1.5%, 2%, 2.5%, 3%, 3.5%,4%, 4.5% or more) of all cells at a radial distance of about 400 μmextending from a site of injection to be mitotically arrested cellswithin about 24 hours of the injection when the composition (such as apharmaceutical composition) is injected into a tumor tissue (for examplewhen injected at the paclitaxel amount of about 12 (or 4 mg/ml) into apancreatic MIA PaCa-2 xenograft tumor). In some embodiments, thecomposition (such as a pharmaceutical composition) is suitable formedical use if it allows more than about 1.5% (such as more than aboutany one of 0.5%, 0.8%, 1%, 1.5%, 2%, 2.5%, 3% or more) of all cells at aradial distance of about 400 μm extending from a site of injection to bemitotically arrested cells within about 24 hours of the injection whenthe composition (such as a pharmaceutical composition) is injected intoa tumor tissue (for example when injected at the paclitaxel amount ofabout 12 μg (or 4 mg/ml) into a pancreatic MIA PaCa-2 xenograft tumor).In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for medical use if it allows more than about 1%(such as more than about any one of 0.4%, 0.6%, 0.8%, 1%, 1.5%, 2% ormore) of all cells at a radial distance of about 400 μm extending from asite of injection to be mitotically arrested cells within about 24 hoursof the injection when the composition (such as a pharmaceuticalcomposition) is injected into a tumor tissue (for example when injectedat the paclitaxel amount of about 12 μg (or 4 mg/ml) into a pancreaticMIA PaCa-2 xenograft tumor). In some embodiments, suitability of medicaluse is determined based on two or more of the above-described functionalattributes.

Thus, the present application in one aspect also provides methods ofdetermining distribution of paclitaxel in a tumor tissue. For example,in some embodiments there is provided a method of determiningdistribution of paclitaxel in a tumor tissue upon administration of analbumin-based paclitaxel nanoparticle composition to the tumor tissue,comprising 1) injecting the composition (such as a pharmaceuticalcomposition) directly to the tumor tissue; and 2) assessing the spatialdistribution of paclitaxel within the tumor tissue. In some embodiments,there is provided a method of determining distribution of paclitaxel ina tumor tissue upon administration of an albumin-based paclitaxelnanoparticle composition to the tumor tissue, comprising 1)microinjecting (for example by using a CIVO™ arrayed microinjectiondevice (Presage Biosciences, Seattle, Wash.) the composition (such as apharmaceutical composition) directly to the tumor tissue; and 2)assessing the spatial distribution of paclitaxel within the tumortissue. In some further embodiments, the injection further comprisesinjection of an imaging agent. In some embodiments, the composition(such as a pharmaceutical composition) is co-injected with an imagingagent to determine the location of the injection site. In someembodiments, the composition (such as a pharmaceutical composition) isco-injected with VivoTag680-S to determine the location of the injectionsite.

In some embodiments, there is provided a method of determiningdistribution of paclitaxel in a tumor tissue upon administration of analbumin-based paclitaxel nanoparticle composition to the tumor tissue,comprising 1) microinjecting (for example by using a CIVO™ arrayedmicroinjection device (Presage Biosciences, Seattle, Wash.) thecomposition (such as a pharmaceutical composition) directly to the tumortissue; and 2) staining the tissue with an agent that indicate thepresence of paclitaxel; and 3) determining the radial distance ofpaclitaxel distribution extending from the site of injection. In someembodiments, the agent that indicates the presence of paclitaxel is anindicator of mitotic arrest such as an anti-pHH3 detection agent (forexample an antibody).

In some embodiments, there is provided a method of determiningdistribution of paclitaxel in a tumor tissue upon administration of analbumin-based paclitaxel nanoparticle composition to the tumor tissue,comprising 1) microinjecting (for example by using a CIVO™ arrayedmicroinjection device (Presage Biosciences, Seattle, Wash.) thecomposition (such as a pharmaceutical composition) directly to the tumortissue; and 2) staining the tissue with an agent that indicate mitoticarrest of a cell; and 3) determining the radial distance of paclitaxeldistribution extending from the site of injection.

In some embodiments, there is provided a method of determiningdistribution of paclitaxel in a tumor tissue upon administration of analbumin-based paclitaxel nanoparticle composition to the tumor tissue,comprising 1) microinjecting (for example by using a CIVO™ arrayedmicroinjection device (Presage Biosciences, Seattle, Wash.) thecomposition (such as a pharmaceutical composition) directly to the tumortissue; and 2) staining the tissue with an agent that indicate themitotic arrest of the cell; and 3) determining the fraction of stainedcells a given radial distance extending from the injection site. In someembodiments, the agent that indicates mitotic arrest of a cell is ananti-pHH3 detection agent (for example an antibody).

In some embodiments, there is provided a method of determiningdistribution of paclitaxel in a tumor tissue upon administration of analbumin-based paclitaxel nanoparticle composition to the tumor tissue,comprising 1) microinjecting (for example by using a CIVO™ arrayedmicroinjection device (Presage Biosciences, Seattle, Wash.) thecomposition (such as a pharmaceutical composition) directly to the tumortissue; and 2) staining the tissue with an agent that indicate themitotic arrest of the cell; and 3) determining the fraction of cells inmitotic arrest at a given radial distance extending from the injectionsite. In some embodiments, the agent that indicates mitotic arrest of acell is an anti-pHH3 detection agent (for example an antibody).

Weight Ratio of the Total Albumin to the Total Paclitaxel in theComposition

The albumin-based paclitaxel nanoparticle compositions assessed bymethods descried herein in some embodiments have a specific albumin topaclitaxel ratio. For example, in some embodiments, the weight ratio ofthe total albumin to the total paclitaxel in the composition (such as apharmaceutical composition) is about 3:1 to about 7.9:1 or about 10:1 toabout 17:1. In some embodiments, the weight ratio of the total albuminto the total paclitaxel in the composition (such as a pharmaceuticalcomposition) is about 3:1 to about 7.9:1, which including for exampleabout 4:1 to about 7:1, about 5:1 to about 7:1, about 6:1 to about 7:1,about 7:1 to about 7.5:1, and about 7.5:1 to about 7.9:1. In someembodiments, the weight ratio of the total albumin to the totalpaclitaxel in the composition (such as a pharmaceutical composition) isabout 10:1 to about 17:1, which include for example, about 10:1 to about15:1, about 10:1 to about 12:1, about 10:1 to about 11:1, or about 10:1to about 10.5:1.

In some embodiments, the weight ratio of the total albumin to the totalpaclitaxel in the composition (such as a pharmaceutical composition) isabout 8:1 to about 10:1. In some embodiments, the weight ratio of thetotal albumin to the total paclitaxel in the composition (such as apharmaceutical composition) is about 9:1.

In some embodiments, the weight ratio of the total albumin to the totalpaclitaxel in the composition (such as a pharmaceutical composition) isabout any of 1:1, 2:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1, 5.5:1, 6:1, 6.5:1,7:1, 7.5:1, 8:1, 8.5:1, 9:1, 9.5:1, 10:1, 10.5:1, 11:1, 12:1, 13:1,14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, theweight ratio of the total albumin to the total paclitaxel in thecomposition (such as a pharmaceutical composition) is any of about 1:1to about 2:1, about 2:1 to about 3:1, about 3:1 to about 3.5:1, about3.5:1 to about 4:1, about 4:1 to about 4.5:1, about 4.5:1 to about 5:1,about 5:1 to about 5.5:1, about 5.5:1 to about 6:1, about 6:1 to about6.5:1, about 6.5:1 to about 7:1, about 7:1 to about 7.5:1, about 7.5:1to about 8:1, about 8:1 to about 8.5:1, about 8.5:1 to about 9:1, about9:1 to about 9.5:1, about 9.5:1 to about 10:1, about 10:1 to about10.5:1, about 10.5:1 to about 11:1, about 11:1 to about 12:1, about 12:1to about 13:1, about 13:1 to about 14:1, about 14:1 to about 15:1, about15:1 to about 16:1, about 16:1 to about 17:1, about 17:1 to about 18:1,about 18:1 to about 19:1, about 19:1 to about 20:1, about 1:1 to about3:1, about 3:1 to about 4:1, about 4:1 to about 5:1, about 5:1 to about6:1, about 6:1 to about 7:1, about 7:1 to about 8:1, about 8:1 to about9:1, about 9:1 to about 10:1, about 10:1 to about 11:1, about 11:1 toabout 13:1, about 13:1 to about 15:1, about 15:1 to about 17:1, about17:1 to about 19:1, about 3:1 to about 5:1, about 5:1 to about 7:1,about 7:1 to about 9:1, about 9:1 to about 11:1, about 2:1 to about 4:1,about 4:1 to about 6:1, about 6:1 to about 8:1, about 8:1 to about 12:1,about 12:1 to about 14:1, about 14:1 to about 16:1, about 16:1 to about18:1, about 3:1 to about 7:1, about 7:1 to about 11:1, about 11:1 toabout 15:1, about 15:1 to about 19:1, about 4:1 to about 8:1, about 8:1to about 12:1, about 12:1 to about 16:1, or about 16:1 to about 20:1.

In some embodiments, to determine the weight ratio of the total albuminto the total paclitaxel in the composition (such as a pharmaceuticalcomposition), the amount of the albumin on the nanoparticle, the amountof the albumin not on the nanoparticle (e.g., in the non-nanoparticleportion, or free in solution in the composition (such as apharmaceutical composition)), the amount of the paclitaxel in thenanoparticle, and the amount of the paclitaxel not associated with thenanoparticle (e.g., in the non-nanoparticle portion, or free in solutionin the composition (such as a pharmaceutical composition)) are needed.As discussed previously, the amount of the albumin on the nanoparticlescan be determined by, for example, chromatography, such assize-exclusion chromatography, or spectrophotometric measurementsfollowing isolation of the nanoparticles in the pharmaceuticalcomposition. In some embodiments, for example, followingultracentrifugation to pellet the nanoparticles, the amount of albuminin the resulting supernatant can be determined by similar methodsdiscussed above for determining the amount of the albumin on thenanoparticles. As discussed previously, the amount of the paclitaxel inthe nanoparticles can be determined by, for example, chromatography,such as RP-HPLC, spectrophotometric measurements, or mass spectrometricmeasurements. In some embodiments, for example, followingultracentrifugation to pellet the nanoparticles, the amount ofpaclitaxel in the resulting supernatant can be determined by similarmethods discussed above for determining the amount of paclitaxel not onthe nanoparticles.

In some embodiments, to determine the weight ratio of the total albuminto the total paclitaxel in the composition (such as a pharmaceuticalcomposition), the amount of the total albumin in the composition (suchas a pharmaceutical composition) and the amount of the total paclitaxelin the composition (such as a pharmaceutical composition) are needed.The amount of the total albumin, for example, can be measured using themethods discussed above using a sample that contains all albumins fromthe composition (such as a pharmaceutical composition). The amount ofthe total paclitaxel, for example, can be measured using the methodsdiscussed above using a sample that contains all paclitaxel from thecomposition (such as a pharmaceutical composition).

The weight ratio of the total albumin to the total paclitaxel in thecomposition (such as a pharmaceutical composition) can be determined,for example, by calculating the ratio of the amount of the total albuminin the composition (such as a pharmaceutical composition) and the amountof the total paclitaxel in the composition (such as a pharmaceuticalcomposition).

Other Components in the Albumin-Based Paclitaxel NanoparticleComposition

The compositions (such as pharmaceutical compositions) described hereinmay also include an antimicrobial agent (e.g., an agent in addition tothe paclitaxel) in an amount sufficient to significantly inhibit (e.g.,delay, reduce, slow, and/or prevent) microbial growth in the composition(such as a pharmaceutical composition) for use in the methods oftreatment, methods of administration, and dosage regimens describedherein. Exemplary microbial agents and variations for the use ofmicrobial agents are disclosed in US 2007/0117744A1 (such as thosedescribed in paragraphs [0036] to [0058] therein), the content of whichis hereby incorporated by reference in its entirety. In someembodiments, the antimicrobial agent is a chelating agent, such as EDTA,edetate, citrate, pentetate, tromethamine, sorbate, ascorbate,derivatives thereof, or mixtures thereof. In some embodiments, theantimicrobial agent is a polydentate chelating agent. In someembodiments, the antimicrobial agent is a non-chelating agent, such asany of sulfites, benzoic acid, benzyl alcohol, chlorobutanol, andparaben. In some embodiments, an antimicrobial other than the taxanediscussed above is not contained or used in the methods of treatment,methods of administration, and dosage regimens described herein.

In some embodiments, the compositions (such as pharmaceuticalcompositions) described herein include a sugar. Exemplary sugars andvariations for the use of sugars are disclosed in US 2007/0117744A1(such as those described in paragraphs [0084] to [0090] therein), thecontent of which is hereby incorporated by reference in its entirety. Insome embodiments, the sugar serves as a reconstitution enhancer whichcauses a lyophilized composition to dissolve or suspend in water and/oraqueous solution more quickly than the lyophilized composition woulddissolve without the sugar. In some embodiments, the composition (suchas a pharmaceutical composition) is a liquid (e.g., aqueous) compositionobtained by reconstituting or resuspending a dry composition. In someembodiments, the concentration of sugar in the composition (such as apharmaceutical composition) is greater than about 50 mg/ml. In someembodiments, the sugar is in an amount that is effective to increase thestability of the paclitaxel in the composition (such as a pharmaceuticalcomposition) as compared to a composition (such as a pharmaceuticalcomposition) without the sugar. In some embodiments, the sugar is in anamount that is effective to improve filterability of the composition(such as a pharmaceutical composition) as compared to a composition(such as a pharmaceutical composition) without the sugar.

The sugar-containing compositions (such as pharmaceutical compositions)described herein may further comprise one or more antimicrobial agents,such as the antimicrobial agents described herein or in US2007/0117744A1. In addition to one or more sugars, other reconstitutionenhancers (such as those described in US 2005/0152979A1, which is herebyincorporated by reference in its entirety) can also be added to thecompositions (such as pharmaceutical compositions).

The compositions (such as pharmaceutical compositions) described hereinmay be used in pharmaceutical compositions or formulations, by combiningthe pharmaceutical composition(s) described with a pharmaceuticallyacceptable carrier, excipients, stabilizing agents and/or other agents,which are known in the art, for use in the methods of treatment, methodsof administration, and dosage regimens described herein.

To increase stability by increasing the negative zeta-potential ofnanoparticles, certain negatively charged components may be added. Suchnegatively charged components include, but are not limited to bilesalts, bile acids, glycocholic acid, cholic acid, chenodeoxycholic acid,taurocholic acid, glycochenodeoxycholic acid, taurochenodeoxycholicacid, litocholic acid, ursodeoxycholic acid, dehydrocholic acid, andothers; phospholipids including lecithin (egg yolk) based phospholipidswhich include the following phosphatidylcholines:palmitoyloleoylphosphatidylcholine,palmitoyllinoleoylphosphatidylcholine,stearoyllinoleoylphosphatidylcholine, stearoyloleoylphosphatidylcholine,stearoylarachidoylphosphatidylcholine, anddipalmitoylphosphatidylcholine. Other phospholipids includingL-α-dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine(DOPC), di stearoylphosphatidylcholine (DSPC), hydrogenated soyphosphatidylcholine (HSPC), and other related compounds. Negativelycharged surfactants or emulsifiers are also suitable as additives, e.g.,sodium cholesteryl sulfate and the like.

Suitable pharmaceutical carriers include sterile water; saline,dextrose; dextrose in water or saline; condensation products of castoroil and ethylene oxide combining about 30 to about 35 moles of ethyleneoxide per mole of castor oil; liquid acid; lower alkanols; oils such ascorn oil; peanut oil, sesame oil and the like, with emulsifiers such asmono- or di-glyceride of a fatty acid, or a phosphatide, e.g., lecithin,and the like; glycols; polyalkylene glycols; aqueous media in thepresence of a suspending agent, for example, sodiumcarboxymethylcellulose; sodium alginate; poly(vinylpyrolidone); and thelike, alone, or with suitable dispensing agents such as lecithin;polyoxyethylene stearate; and the like. The carrier may also containadjuvants such as preserving stabilizing, wetting, emulsifying agentsand the like together with the penetration enhancer. The final form maybe sterile and may also be able to pass readily through an injectiondevice such as a hollow needle. The proper viscosity may be achieved andmaintained by the proper choice of solvents or excipients. Moreover, theuse of molecular or particulate coatings such as lecithin, the properselection of particle size in dispersions, or the use of materials withsurfactant properties may be utilized.

The pharmaceutical compositions described herein may include otheragents, excipients, or stabilizers to improve properties of thecomposition (such as a pharmaceutical composition). Examples of suitableexcipients and diluents include, but are not limited to, lactose,dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calciumphosphate, alginates, tragacanth, gelatin, calcium silicate,microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water,saline solution, syrup, methylcellulose, methyl- andpropylhydroxybenzoates, talc, magnesium stearate and mineral oil. Theformulations can additionally include lubricating agents, wettingagents, emulsifying and suspending agents, preserving agents, sweeteningagents or flavoring agents. Examples of emulsifying agents includetocopherol esters such as tocopheryl polyethylene glycol succinate andthe like, Pluronic®, emulsifiers based on polyoxy ethylene compounds,Span 80 and related compounds and other emulsifiers known in the art andapproved for use in animals or human dosage forms. The compositions(such as pharmaceutical compositions) can be formulated so as to providerapid, sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures well known in theart.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is formulated to have a pH in the range of about 4.5 toabout 9.0, including for example pH ranges of any one of about 5.0 toabout 8.0, about 6.5 to about 7.5, or about 6.5 to about 7.0. In someembodiments, the pH of the composition (such as a pharmaceuticalcomposition) is formulated to no less than about 6, including forexample no less than about any one of 6.5, 7, or 8 (e.g., about 8). Thecomposition (such as a pharmaceutical composition) can also be made tobe isotonic with blood by the addition of a suitable tonicity modifier,such as glycerol.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is suitable for administration to a human. In someembodiments, the composition (such as a pharmaceutical composition) issuitable for administration to a human by parenteral administration.Formulations suitable for parenteral administration include aqueous andnon-aqueous, isotonic sterile injection solutions, which can containanti-oxidants, buffers, bacteriostats, and solutes that render theformulation compatible with the blood of the intended recipient, andaqueous and non-aqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizing agents, andpreservatives. The formulations can be presented in unit-dose ormulti-dose sealed containers, such as ampules and vials, and can bestored in a freeze-dried (lyophilized) condition requiring only theaddition of the sterile liquid excipient methods of treatment, methodsof administration, and dosage regimens described herein (e.g., water)for injection, immediately prior to use. Extemporaneous injectionsolutions and suspensions can be prepared from sterile powders,granules, and tablets of the kind previously described. Injectableformulations are preferred. In some embodiments, the composition (suchas a pharmaceutical composition) is contained in a single-use vial, suchas a single-use sealed vial. In some embodiments, each single-use vialcontains about 100 mg paclitaxel. In some embodiments, the single-usevial contains about 900 mg albumin. In some embodiments, the composition(such as a pharmaceutical composition) is contained in a multi-use vial.In some embodiments, the composition (such as a pharmaceuticalcomposition) is contained in bulk in a container.

Also provided are unit dosage forms comprising the compositions (such aspharmaceutical compositions) and formulations described herein. Theseunit dosage forms can be stored in a suitable packaging in single ormultiple unit dosages and may also be further sterilized and sealed. Insome embodiments, the composition (such as a pharmaceutical composition)(such as a pharmaceutical composition) also includes one or more othercompounds (or pharmaceutically acceptable salts thereof) that are usefulfor treating cancer. In various variations, the amount of paclitaxel inthe composition (such as a pharmaceutical composition) is included inany one of the following ranges: about 5 mg to about 50 mg, about 20 mgto about 50 mg, about 50 mg to about 100 mg, about 100 mg to about 125mg, about 125 mg to about 150 mg, about 150 mg to about 175 mg, about175 mg to about 200 mg, about 200 mg to about 225 mg, about 225 mg toabout 250 mg, about 250 mg to about 300 mg, about 300 mg to about 350mg, about 350 mg to about 400 mg, about 400 mg to about 450 mg, or about450 mg to about 500 mg. In some embodiments, the amount of paclitaxel inthe composition (such as a pharmaceutical composition) (e.g., a dosageor unit dosage form) is in the range of about 5 mg to about 500 mg, suchas about 30 mg to about 300 mg or about 50 mg to about 200 mg. In someembodiments, the carrier is suitable for parental administration (e.g.,intravenous administration). In some embodiments, the paclitaxel is theonly pharmaceutically active agent for the treatment of cancer that iscontained in the composition (such as a pharmaceutical composition).

In some embodiments, there is provided a dosage form (e.g., a unitdosage form) for the treatment of cancer comprising any one of thecompositions (such as pharmaceutical compositions) described herein. Insome embodiments, there are provided articles of manufacture comprisingthe compositions (such as pharmaceutical compositions), formulations,and unit dosages described herein in suitable packaging for use in themethods of treatment, methods of administration, and dosage regimensdescribed herein. Suitable packaging for compositions (such aspharmaceutical compositions) described herein are known in the art, andinclude, for example, vials (such as sealed vials), vessels (such assealed vessels), ampules, bottles, jars, flexible packaging (e.g.,sealed Mylar or plastic bags), and the like. These articles ofmanufacture may further be sterilized and/or sealed.

Methods of Validating and/or Releasing a Commercial Batch ofAlbumin-Based Paclitaxel Nanoparticle Compositions

The methods described herein may be carried out when validating and/orreleasing a commercial batch of an albumin-based paclitaxel nanoparticlecomposition. “Commercial batch” used herein refers to a batch size thatis at least about 20 grams (by amount of paclitaxel). In someembodiments, the batch size is at least about 30 grams, 40 grams, 50grams, 60 grams, 70 grams, 80 grams, 90 grams, 100 grams, 150 grams, 200grams, 250 grams, 300 grams, 350 grams, 400 grams, 450 grams, 500 grams,550 grams, 600 grams, 650 grams, 700 grams, 750 grams, 800 grams, 850grams, 900 grams, 1000 grams, 1500 grams, 2000 grams, 2500 grams, 3000grams, 3500 grams, 4000 grams, 4500 grams, 5000 grams, or 10,000 grams(by amount of paclitaxel). In some embodiments, the commercial batchcomprises a plurality of vials comprising any of the compositions (suchas pharmaceutical compositions) described herein. In some embodiments,the commercial batch comprises at least about any of 100 vials, 150vials, 200 vials, 100 vials, 150 vials, 200 vials, 250 vials, 300 vials,350 vials, 400 vials, 450 vials, 500 vials, 550 vials, 600 vials, 650vials, 700 vials, 750 vials, 800 vials, 850 vials, 900 vials, 1000vials, 1500 vials, 2000 vials, 2500 vials, 3000 vials, 3500 vials, 4000vials, 4500 vials, 5000 vials, 10000 vials, 12000 vials, 14000 vials,16000 vials, 18000 vials, 20000 vials, 22000 vials, 24000 vials, 26000vials, 28000 vials, 30000 vials, 32000 vials, 34000 vials, 36000 vials,38000 vials, 40000 vials, 42000 vials, 44000 vials, 46000 vials, 48000vials, or 50000 vials. For example, each vial contains about any of 100mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, or1000 mg of the composition (such as a pharmaceutical composition). Insome embodiments, the pharmaceutical composition in the commercial batchis a liquid suspension. In some embodiments, the pharmaceuticalcomposition in the commercial batch is a lyophilized powder.

The present application thus in some embodiments provides a method ofvalidating and/or releasing a commercial batch of a pharmaceuticalcomposition for medical use in a human individual, wherein thepharmaceutical composition comprise nanoparticles comprising paclitaxelcoated with albumin and a non-nanoparticle portion comprising albuminand paclitaxel, the method comprises a) obtaining a sample from thecommercial batch; and 2) assessing suitability of the pharmaceuticalcomposition for medical use according to any one of the methodsdescribed herein. In some embodiments, at least about 2 samples, 3samples, 4 samples, 5 samples, 6 samples, 10 samples, 20 samples, 30samples, 40 samples, 50 samples, 60 samples, 70 samples, 80 samples, 90samples, 100 samples, or more samples are obtained from the commercialbatch and subject to assessment. In some embodiments, the amount of thesample (i.e., the amount of the pharmaceutical composition taken fromthe commercial batch) is about any of 10-20 μg, 20-50 μg, 50-100 μg,100-200 μg, 200-500 μg, 500-1000 μg, 1000 μg, 2000 μg, 3000 μg, 4000 μg,5000 μg, or greater than 5000 μg. In some embodiments, the sample is thepharmaceutical composition in a vial. In some embodiments, the sample isobtained from the commercial batch prior to lyophilization of thecomposition (such as a pharmaceutical composition). In some embodiments,the sample is obtained from the commercial batch after lyophilization ofthe composition (such as a pharmaceutical composition). In someembodiments, the sample is obtained from the commercial batch afterreconstitution.

Thus, for example, the present application in some embodiments providesa method of validating and/or releasing a commercial batch of apharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: a) obtaining asample from the commercial batch; and b) determining the percentage ofalbumin polymers among the albumin on the nanoparticles in the sample,wherein a percentage of albumin polymer among the albumin on thenanoparticles being about 15% to about 40% (such as about 15% to about20%, about 20% to about 24.5%, about 24.5% to about 30%, about 30% toabout 35%, or about 35% to about 40%) is indicative of suitability ofthe commercial batch for medical use and/or release.

In some embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the pharmaceutical compositioncomprises nanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: a) obtaining a sample from the commercial batch; and b)determining the percentage of albumin monomers among the albumin on thenanoparticles in the sample, wherein a percentage of albumin monomersamong the albumin on the nanoparticles being about 40% to about 60%(such as about 40% to about 55%, about 40% to about 54%, about 40% toabout 53%, about 40% to about 52%, about 40% to about 50%, about 40% toabout 48%, or about 40% to about 46%) is indicative of suitability ofthe commercial batch for medical use and/or release.

In some embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the pharmaceutical compositioncomprises nanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: a) obtaining a sample from the commercial batch; and b)determining the weight percentage of the albumin in the nanoparticles,wherein a weight percentage of the albumin in the nanoparticles beingabout 15% to about 30% (such as about 20% to about 25%, about 15% toabout 24%, or about 15% to about 20%) is indicative of suitability ofthe commercial batch for medical use and/or release.

In some embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the pharmaceutical compositioncomprises nanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: a) obtaining a sample from the commercial batch; and b)determining the weight ratio of albumin to paclitaxel in thenanoparticles, wherein an albumin to paclitaxel ratio of about 1:2 toabout 1:6 in the nanoparticles is indicative of suitability of thecommercial batch for medical use and/or release.

In some embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the pharmaceutical compositioncomprises nanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: a) obtaining a sample from the commercial batch; and b)determining the morphology of the nanoparticles under cryo-TEM, whereinan irregular shape of the nanoparticles is indicative of suitability ofthe commercial batch for medical use and/or release.

In some embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the pharmaceutical compositioncomprises nanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: a) obtaining a sample from the commercial batch; and b)determining the thickness of the albumin coating of the nanoparticlesunder cryo-TEM, wherein a thickness of about 5-7 nm (such as about 6 nm)is indicative of suitability of the commercial batch for medical useand/or release.

In some embodiments, there is provided a method of validating and/orrelease a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the pharmaceutical compositioncomprises nanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: a) obtaining a sample from the commercial batch; and b)determining the distribution of paclitaxel in a tumor tissue uponinjection of the pharmaceutical composition directly into the tumortissue; wherein an enhanced paclitaxel tumor distribution is indicativeof suitability of the commercial batch for medical use and/or release.

In some embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the pharmaceutical compositioncomprises nanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: a) obtaining a sample from the commercial batch; and b)determining the solubility, paclitaxel crystallinity, and a recoveryfollowing a 0.2 micron filtration of the pharmaceutical composition,wherein a solubility of about 50 μg/ml to about 100 μg/ml in a 5% humanalbumin solution, a non-crystalline state of the paclitaxel, and arecovery date of at least about 80% is indicative of suitability of thecommercial batch for medical use and/or release.

In some embodiments, the method comprises various combinations of thedetermination steps described above. In some embodiments, the methodcomprises at least 2, 3, 4, 5, 6, 7, or 8 determination steps describedabove. In some embodiments, the method further comprises determining atleast one (such as at least any of 2, 3, 4, 5, 6, 7, 8, 9, or 10) of thefollowing characteristics of the sample: 1) binding affinity of albuminto paclitaxel in the composition (such as a pharmaceutical composition)(for example by equilibrium dialysis, FTIR, NMR, or a combinationthereof); 2) surface-to-volume ratio; 3) percentage of albumin dimersand/or oligomers among the albumin on the nanoparticles; 4) distributionof the total paclitaxel and/or the total albumin between thenanoparticles and the non-nanoparticle portion; 5) oligomeric status ofthe total albumin in the composition (such as a pharmaceuticalcomposition); 6) particle size of the nanoparticles, including averageparticle size, polydispersity, and/or size distribution; 7) surfacepotential; 8) in vitro release kinetics; 9) physical stability; and 10)paclitaxel tumor distribution in vivo. In some embodiments, at least one(such as at least any of 2, 3, 4, 5, 6, 7, 8, or 9) of the determinationsteps are carried out upon reconstitution of the sample. In someembodiments, at least one (such as at least any of 2, 3, 4, 5, 6, 7, 8,or 9) of the determination steps are carried out upon storage of thesample. In some embodiments, at least one of the determination steps arecarried out after storage of the sample for at least about any of 6hours, 12 hours, 24 hours, 36 hours, 48 hours, or 72 hours. In someembodiments, at least one of the determination steps are carried outafter storage of the sample at room temperature, under refrigeratedcondition, or at about 40° C.

In some embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the pharmaceutical compositioncomprises nanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: determining the solubility, paclitaxel crystallinity, andpaclitaxel recovery following a 0.2 micron filtration of thepharmaceutical composition, determining the percentage of albuminmonomers, dimers, oligomers, or polymers among the total albumin in thepharmaceutical composition, determining the percentage of the paclitaxelin the nanoparticles among the total paclitaxel in the pharmaceuticalcomposition (for example by reversed-phase HPLC), determining thepercentage of the albumin that is in the non-nanoparticle portion amongthe total albumin in the pharmaceutical composition (for example bysize-exclusion chromatography), determining the particle size of thenanoparticles (for example by dynamic light scattering) and/or sizedistribution of the nanoparticles, and determining the stability of thepharmaceutical composition (including determining stability afterstorage). In some embodiments, the solubility, paclitaxel crystallinity,and/or paclitaxel recovery are determined after storage (for exampleafter storage for at least about any of 6 hours, 12 hours, 18 hours, 24hours, 36 hours, 48 hours, or 72 hours, such as at room temperature,under refrigerated condition, or at about 40° C.). In some embodiments,the paclitaxel crystallinity is determined by X-ray diffraction and/orpolarized light microscopy. In some embodiments, the method furthercomprises determining binding affinity of albumin to paclitaxel in thecomposition (such as a pharmaceutical composition) (for example byequilibrium dialysis, FTIR, NMR, or a combination thereof). In someembodiments, the method further comprises determining tumor distributionof paclitaxel upon administration in vivo (for example by determiningtumor distribution of paclitaxel upon injection of the pharmaceuticalcomposition directly into the tumor tissue).

In some embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein the nanoparticles having less than about 52% of the albumin onthe nanoparticles in the form of monomers is indicative of suitabilityof the commercial batch for medical use and/or release. In someembodiments, there is provided a method of validating and/or releasing acommercial batch of a pharmaceutical composition for medical use in ahuman individual, wherein the commercial batch of a pharmaceuticalcomposition comprising a pharmaceutical composition comprising: a)nanoparticles comprising paclitaxel coated with albumin, and b) anon-nanoparticle portion comprising albumin and paclitaxel, wherein thenanoparticles having more than about 35% of albumin on the nanoparticlesin the forms of polymers and oligomers is indicative of suitability ofthe commercial batch for medical use and/or release. In someembodiments, there is provided a method of validating and/or releasing acommercial batch of a pharmaceutical composition for medical use in ahuman individual, wherein the commercial batch of a pharmaceuticalcomposition comprising a pharmaceutical composition comprising: a)nanoparticles comprising paclitaxel coated with albumin, and b) anon-nanoparticle portion comprising albumin and paclitaxel, wherein thenanoparticles having less than about 54% of the albumin on thenanoparticles in the form of monomers, and more than about 11% of thealbumin in the nanoparticles in the form of polymers is indicative ofsuitability of the commercial batch for medical use and/or release. Insome embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein the nanoparticles having less than about 55% of the albumin onthe nanoparticles in the form of monomers, and more than about 18% ofthe albumin in the nanoparticles in the form of polymers is indicativeof suitability of the commercial batch for medical use and/or release.In some embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein the nanoparticles having a ratio of albumin on the nanoparticlesin the form of polymers and oligomers divided by the albumin on thenanoparticles in the form of monomers of more than about 62% isindicative of suitability of the commercial batch for medical use and/orrelease. In some embodiments, there is provided a method of validatingand/or releasing a commercial batch of a pharmaceutical composition formedical use in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein the nanoparticles having a weight percentage of the albumin inthe nanoparticles of about 15% to about 30% (such as about 20% to about25%, about 15% to about 24%, or about 15% to about 20%) is indicative ofsuitability of the commercial batch for medical use and/or release. Insome embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein the nanoparticles having a weight ratio of the albumin to thepaclitaxel in the nanoparticles of about 1:2 to about 1:6 is indicativeof suitability of the commercial batch for medical use and/or release.In some embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein the nanoparticles in the pharmaceutical composition having anirregular shape is indicative of suitability of the commercial batch formedical use and/or release. In some embodiments, there is provided amethod of validating and/or releasing a commercial batch of apharmaceutical composition for medical use in a human individual,wherein the commercial batch of a pharmaceutical composition comprisinga pharmaceutical composition comprising: a) nanoparticles comprisingpaclitaxel coated with albumin, and b) a non-nanoparticle portioncomprising albumin and paclitaxel, wherein a thickness of the albumincoating on the nanoparticles of about 5-7 nm as measured by cryo-TEM isindicative of suitability of the commercial batch for medical use and/orrelease. In some embodiments, there is provided a method of validatingand/or releasing a commercial batch of a pharmaceutical composition formedical use in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein a weight ratio of the total albumin to the total paclitaxel inthe pharmaceutical composition of about 3:1 to about 7.9:1 or about 10:1to about 17:1 is indicative of suitability of the commercial batch formedical use and/or release. In some embodiments, there is provided amethod of validating and/or releasing a commercial batch of apharmaceutical composition for medical use in a human individual,wherein the commercial batch of a pharmaceutical composition comprisinga pharmaceutical composition comprising: a) nanoparticles comprisingpaclitaxel coated with albumin, and b) a non-nanoparticle portioncomprising albumin and paclitaxel, wherein a solubility of about 50μg/ml to about 100 μg/ml (including for example any of about 50 μg/ml toabout 60 μg/ml, about 60 μg/ml to about 70 μg/ml, about 70 μg/ml toabout 75 μg/ml, about 75 μg/ml to about 80 μg/ml, about 80 μg/ml toabout 90 μg/ml, or about 90 μg/ml to about 100 μg/ml) when diluted in a5% human albumin solution is indicative of suitability of the commercialbatch for medical use and/or release. In some embodiments, there isprovided a method of validating and/or releasing a commercial batch of apharmaceutical composition for medical use in a human individual,wherein the commercial batch of a pharmaceutical composition comprisinga pharmaceutical composition comprising: a) nanoparticles comprisingpaclitaxel coated with albumin, and b) a non-nanoparticle portioncomprising albumin and paclitaxel, wherein a paclitaxel recovery of atleast about 80% (including for example at least about any of 85%, 90%,95%, 98%, or 99%) following a 0.2 micron filtration is indicative ofsuitability of the commercial batch for medical use and/or release. Insome embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein an average particle size of the nanoparticles in thepharmaceutical composition of less than about 200 nm (including forexample about 100 to about 160 nm) is indicative of suitability of thecommercial batch for medical use and/or release. In some embodiments,there is provided a method of validating and/or releasing a commercialbatch of a pharmaceutical composition for medical use in a humanindividual, wherein the commercial batch of a pharmaceutical compositioncomprising a pharmaceutical composition comprising: a) nanoparticlescomprising paclitaxel coated with albumin, and b) a non-nanoparticleportion comprising albumin and paclitaxel, wherein the nanoparticles inthe pharmaceutical composition having a zeta-potential of about −20 mVto about −35 mV is indicative of suitability of the commercial batch formedical use and/or release. In some embodiments, there is provided amethod of validating and/or releasing a commercial batch of apharmaceutical composition for medical use in a human individual,wherein the commercial batch of a pharmaceutical composition comprisinga pharmaceutical composition comprising: a) nanoparticles comprisingpaclitaxel coated with albumin, and b) a non-nanoparticle portioncomprising albumin and paclitaxel, wherein the nanoparticles in thepharmaceutical composition having a polydispersity index of less thanabout 0.3 (including for example less than about 0.06 or about any of0.05-0.09, 0.09-0.13, 0.13-0.15, 0.15-0.2, or 0.2-0.3) is indicative ofsuitability of the commercial batch for medical use and/or release. Insome embodiments, there is provided a method of validating and/orreleasing a commercial batch of a pharmaceutical composition for medicaluse in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein the nanoparticles in the pharmaceutical composition having aspan of size distribution ((Dv₉₀−Dv₁₀)/Dv₅₀) of about 0.8 to about 1.5(including for example about any of 0.8-0.9, 0.9-1, 1-1.1, 1.1-1.2,1.2-1.3, 1.3-1.4, or 1.4-1.5) is indicative of suitability of thecommercial batch for medical use and/or release. In some embodiments,there is provided a method of validating and/or releasing a commercialbatch of a pharmaceutical composition for medical use in a humanindividual, wherein the commercial batch of a pharmaceutical compositioncomprising a pharmaceutical composition comprising: a) nanoparticlescomprising paclitaxel coated with albumin, and b) a non-nanoparticleportion comprising albumin and paclitaxel, wherein the paclitaxel in thepharmaceutical composition being non-crystalline (including for examplenon-crystalline after storage for about 24 hours at about 4° C.) isindicative of suitability of the commercial batch for medical use and/orrelease. In some embodiments, there is provided a method of validatingand/or releasing a commercial batch of a pharmaceutical composition formedical use in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein the pharmaceutical composition upon tumor injection allowingpaclitaxel to spread radially for a distance of greater than (forexample more than about any of 1.1×, 1.2×, 1.3×, 1.4×, 1.5×, 1.6×, 1.7×,1.8×, 2×, 3×, 4×, 5×, 6×, 7×, 8×, or more of) that of a solvent-basedpaclitaxel formulation (such as the solvent-based paclitaxel formulationsold under the trademark TAXOL®) under the same assay conditions isindicative of suitability of the commercial batch for medical use and/orrelease. In some embodiments, there is provided a method of validatingand/or releasing a commercial batch of a pharmaceutical composition formedical use in a human individual, wherein the commercial batch of apharmaceutical composition comprising a pharmaceutical compositioncomprising: a) nanoparticles comprising paclitaxel coated with albumin,and b) a non-nanoparticle portion comprising albumin and paclitaxel,wherein the pharmaceutical composition upon tumor injection allowingpaclitaxel to spread radically for more than about 700 μm (such as morethan about any of 700 μm, 800 μm, 900 μm, 1000 μm, 1100 μm or 1200 μm)within about 24 hours after the pharmaceutical composition is injectedinto a tumor tissue (for example injected at the paclitaxel amount ofabout 12 μg (such as at about 4 mg/ml) into a pancreatic MIA PaCa-2xenograft tumor) is indicative of suitability of the commercial batchfor medical use and/or release.

Also provided are commercial batches released by following any one ofthe methods described herein.

Further provided are kits for use in any one of the methods ofassessment and commercial batch release described herein.

Kits Comprising the Albumin-Based Paclitaxel Nanoparticle Compositions

Once determined to be suitable for medical use, the pharmaceuticalcompositions can be included in kits comprising the compositions (suchas pharmaceutical compositions), formulations, unit dosages, andarticles of manufacture for use in the methods of treatment, methods ofadministration, and dosage regimens described herein. Kits describedherein include one or more containers comprising the paclitaxelpharmaceutical compositions (formulations or unit dosage forms and/orarticles of manufacture), and in some embodiments, further compriseinstructions for accessing and/or using in accordance with any of themethods of treatment described herein. In various embodiments, theamount of paclitaxel in the kit is included in any one of the followingranges: about 5 mg to about 20 mg, about 20 mg to about 50 mg, about 50mg to about 100 mg, about 100 mg to about 125 mg, about 125 mg to about150 mg, about 150 mg to about 175 mg, about 175 mg to about 200 mg,about 200 mg to about 225 mg, about 225 mg to about 250 mg, about 250 mgto about 300 mg, about 300 mg to about 350 mg, about 350 mg to about 400mg, about 400 mg to about 450 mg, or about 450 mg to about 500 mg. Insome embodiments, the amount of paclitaxel in the kit is in the range ofabout 5 mg to about 500 mg, such as about 30 mg to about 300 mg or about50 mg to about 200 mg. In some embodiments, the kit includes one or moreother compounds (e.g., one or more compounds other than paclitaxel thatare useful for cancer).

Instructions supplied in the kits described herein are typically writteninstructions on a label or package (e.g., a paper sheet included in thekit), but machine-readable instructions (e.g., instructions carried on amagnetic or optical storage disk) are also acceptable. The instructionsrelating to the use of the pharmaceutical compositions generally includeinformation as to dosage, dosing schedule, and route of administrationfor the intended treatment. The kit may further comprise a descriptionof selecting an individual suitable or treatment.

The present application also provides kits comprising compositions (suchas pharmaceutical compositions) (or unit dosages forms and/or articlesof manufacture) described herein and may further comprise instruction(s)on methods of assessing and/or using the composition (such as apharmaceutical composition), such as uses further described herein. Insome embodiments, the kit described herein comprises the packagingdescribed above. In other variations, the kit described herein comprisesthe packaging described above and a second packaging comprising abuffer. It may further include other materials desirable from acommercial and user standpoint, including other buffers, diluents,filters, needles, syringes, and packages with instructions forperforming any methods described herein.

For combination therapies described herein, the kit may containinstructions for administering the first and second therapiessimultaneously and/or sequentially for the effective treatment ofcancer. The first and second therapies can be present in separatecontainers or in a single container. It is understood that the kit maycomprise one distinct composition or two or more compositions (such aspharmaceutical compositions) wherein one composition comprises a firsttherapy and one composition comprises a second therapy.

Kits may also be provided that contain sufficient dosages of thepaclitaxel as disclosed herein to provide effective treatment for anindividual for an extended period, such as any one of a week, 2 weeks, 3weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 6months, 7 months, 8 months, 9 months or more. Kits may also includemultiple unit doses of the paclitaxel, compositions (such aspharmaceutical compositions), and formulations described herein andinstructions for use and packaged in quantities sufficient for storageand use in pharmacies, for example, hospital pharmacies and compoundingpharmacies. In some embodiments, the kit comprises a dry (e.g.,lyophilized) composition that can be reconstituted, resuspended, orrehydrated to form generally a stable aqueous suspension ofnanoparticles comprising paclitaxel and albumin.

The kits described herein are in suitable packaging. Suitable packaginginclude, but is not limited to, vials, bottles, jars, flexible packaging(e.g., sealed Mylar or plastic bags), and the like. Kits may optionallyprovide additional components such as buffers and interpretativeinformation.

Methods of Making the Pharmaceutical Compositions

The present application also provides methods of making the paclitaxelpharmaceutical compositions described herein. Nanoparticles containingpoorly water soluble pharmaceutical agents and carrier proteins (e.g.,albumin) can be prepared under conditions of high shear forces (e.g.,sonication, high pressure homogenization, or the like). These methodsare disclosed in, for example, U.S. Pat. Nos. 5,916,596; 6,096,331;6,749,868; 6,537,579; and PCT Application Pub. Nos. WO98/14174;WO99/00113; WO07/027941; and WO07/027819. The contents of thesepublications, particularly with respect to the method of makingcompositions (such as pharmaceutical compositions) containing carrierproteins, are hereby incorporated by reference in their entireties.

Generally, to make the paclitaxel pharmaceutical compositions describedherein, paclitaxel is dissolved in an organic solvent. Suitable organicsolvents include, for example, ketones, esters, ethers, chlorinatedsolvents, and other solvents known in the art. For example, the organicsolvent can be methylene chloride/ethanol, chloroform/ethanol, orchloroform/t-butanol (for example with a ratio of about any one of 1:9,1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1,8:1, or 9:1 or with a ratio of about any one of 3:7, 5:7, 4:6, 5:5, 6:5,8:5, 9:5, 9.5:5, 5:3, 7:3, 6:4, or 9.5:0.5). Albumin (such asrecombinant albumin, for example recombinant albumin sold under thetrademark NOVOZYME™ or recombinant albumin sold under the trademarkINTRIVIA™ recombinant albumin disclosed herein) is dissolved in waterand combined with the paclitaxel solution. The mixture is subjected tohigh pressure homogenization (e.g., using a high pressure homogenizersold by Avestin, APV Gaulin, or Stansted, or a high pressure homogenizersold under the trademark MICROFLUIDIZER™ such as the high pressurehomogenizer sold under the trademark MICROFLUIDIZER™ Processor M-110EHsold by Microfluidics, or the high pressure homogenizer sold under thetrademark ULTRA-TURRAX®). The emulsion may be cycled through the highpressure homogenizer for between about 2 to about 100 cycles, such asabout 5 to about 50 cycles or about 8 to about 20 cycles (e.g., aboutany one of 8, 10, 12, 14, 16, 18 or 20 cycles). The organic solvent canthen be removed by evaporation utilizing suitable equipment known forthis purpose, including, but not limited to, rotary evaporators, fallingfilm evaporators, wiped film evaporators, spray driers, and the likethat can be operated in batch mode or in continuous operation. Thesolvent may be removed at reduced pressure (such as at about any one of25 mm Hg, 30 mm Hg, 40 mm Hg, 50 mm Hg, 100 mm Hg, 200 mm Hg, or 300 mmHg). The amount of time used to remove the solvent under reducedpressure may be adjusted based on the volume of the formulation. Forexample, for a formulation produced on a 300 mL scale, the solvent canbe removed at about 1 to about 300 mm Hg (e.g., about any one of 5-100mm Hg, 10-50 mm Hg, 20-40 mm Hg, or 25 mm Hg) for about 5 minutes toabout 60 minutes (e.g., about any one of 7 minutes, 8 minutes, 9minutes, 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes, 15minutes, 16 minutes, 18 minutes, 20 minutes, 25 minutes, or 30 minutes).The dispersion obtained can be further lyophilized.

If desired, additional albumin solution may be added to the dispersionto adjust the albumin to paclitaxel ratio, or to adjust theconcentration of paclitaxel in the dispersion. For example, albuminsolution (e.g., 25% w/v) can be added to adjust the albumin topaclitaxel ratio to about any one of 18:1, 15:1 14:1, 13:1, 12:1, 11:1,10:1, 9:1, 8:1, 7.5:1, 7:1, 6:1, 5:1, 4:1, or 3:1. In another example,albumin solution (e.g., 25% w/v) or another solution is added to adjustthe concentration of paclitaxel in the dispersion to about any one of0.5 mg/ml, 1.3 mg/ml, 1.5 mg/ml, 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, 10 mg/ml, 15 mg/ml, 20 mg/ml, 25mg/ml, 30 mg/ml, 40 mg/ml, or 50 mg/ml. The dispersion may be seriallyfiltered through multiple filters, such as a combination of 1.2 μm and0.8/0.2 μm filters; the combination of 1.2 μm, 0.8 μm, 0.45 μm, and 0.22μm filters; or the combination of any other filters known in the art.The dispersion obtained can be further lyophilized. The pharmaceuticalcompositions may be made using a batch process or a continuous process(e.g., the production of a composition (such as a pharmaceuticalcomposition) on a large scale).

If desired, a second therapy (e.g., one or more compounds useful fortreating cancer), an antimicrobial agent, sugar, and/or stabilizingagent can also be included in the composition (such as a pharmaceuticalcomposition). For example, this additional agent can either be admixedwith paclitaxel and/or the albumin during the preparation of thepaclitaxel pharmaceutical composition, or added after the paclitaxelpharmaceutical composition is prepared. In some embodiments, the agentis admixed with the paclitaxel pharmaceutical composition prior tolyophilization. In some embodiments, the agent is added to thelyophilized paclitaxel pharmaceutical composition. In some embodimentswhen the addition of the agent changes the pH of the composition (suchas a pharmaceutical composition), the pH in the composition (such as apharmaceutical composition) are generally (but not necessarily) adjustedto a desired pH. Exemplary pH values of the compositions (such aspharmaceutical compositions) include, for example, in the range of about5 to about 8.5. In some embodiments, the pH of the composition (such asa pharmaceutical composition) is adjusted to no less than about 6,including for example no less than any one of about 6.5, 7, or 8 (e.g.,about 8).

Methods of Treating Diseases

Once determined suitable for medical use by following the methodsdescribed herein, the pharmaceutical compositions may be used to treatdiseases associated with cellular proliferation or hyperproliferation,such as cancers.

Examples of cancers that may be treated by the methods described hereininclude, but are not limited to, breast cancer (such as metastaticbreast cancer), lung cancer (such as non-small cell lung cancer),pancreatic cancer (such as metastatic pancreatic cancer or locallyadvanced unresectable pancreatic cancer), multiple myeloma, renal cellcarcinoma, prostate cancer, melanoma (such as metastatic melanoma),colon cancer, colorectal cancer, ovarian cancer, liver cancer, renalcancer, and gastric cancer. In some embodiments, the cancer is breastcancer after failure of combination chemotherapy for metastatic diseaseor relapse within 6 months of adjuvant chemotherapy. In someembodiments, the prior therapy includes an anthracycline treatment.

Cancers to be treated by compositions (such as pharmaceuticalcompositions) described herein include, but are not limited to,carcinoma, lymphoma, blastoma, sarcoma, and leukemia. Examples ofcancers that can be treated by compositions (such as pharmaceuticalcompositions) described herein include, but are not limited to, squamouscell cancer, lung cancer (including small cell lung cancer, non-smallcell lung cancer, adenocarcinoma of the lung, and squamous carcinoma ofthe lung, including squamous NSCLC), cancer of the peritoneum,hepatocellular cancer, gastric or stomach cancer (includinggastrointestinal cancer), pancreatic cancer (such as advanced pancreaticcancer), glioblastoma, cervical cancer, ovarian cancer, liver cancer(such as hepatocellular carcinoma), bladder cancer, hepatoma, breastcancer, colon cancer, melanoma, endometrial or uterine carcinoma,salivary gland carcinoma, kidney or renal cancer, prostate cancer (suchas advanced prostate cancer), vulval cancer, thyroid cancer, hepaticcarcinoma, head and neck cancer, colorectal cancer, rectal cancer,soft-tissue sarcoma, Kaposi's sarcoma, B-cell lymphoma (including lowgrade/follicular non-Hodgkin's lymphoma (NHL), small lymphocytic (SL)NHL, intermediate grade/follicular NHL, intermediate grade diffuse NHL,high grade immunoblastic NHL, high grade lymphoblastic NHL, high gradesmall non-cleaved cell NHL, bulky disease NHL, mantle cell lymphoma,AIDS-related lymphoma, and Waldenstrom's macroglobulinemia), chroniclymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), myeloma,Hairy cell leukemia, chronic myeloblastic leukemia, and post-transplantlymphoproliferative disorder (PTLD), as well as abnormal vascularproliferation associated with phakomatoses, edema (such as thatassociated with brain tumors), and Meigs' syndrome. In some embodiments,there is provided a method of treating metastatic cancer (that is,cancer that has metastasized from the primary tumor). In someembodiments, there is provided a method of reducing cell proliferationand/or cell migration. In some embodiments, there is provided a methodof treating hyperplasia, for example hyperplasia in the vascular systemthat can result in restenosis or hyperplasia that can result in arterialor venous hypertension.

In some embodiments, there are provided methods of treating cancer atadvanced stage(s). In some embodiments, there are provided methods oftreating breast cancer (which may be HER2 positive or HER2 negative),including, for example, advanced breast cancer, stage IV breast cancer,locally advanced breast cancer, and metastatic breast cancer. In someembodiments, the cancer is lung cancer, including, for example,non-small cell lung cancer (NSCLC, such as advanced NSCLC), small celllung cancer (SCLC, such as advanced SCLC), and advanced solid tumormalignancy in the lung. In some embodiments, the cancer is ovariancancer, head and neck cancer, gastric malignancies, melanoma (includingmetastatic melanoma), colorectal cancer, pancreatic cancer, and solidtumors (such as advanced solid tumors). In some embodiments, the canceris any of (and in some embodiments selected from the group consistingof) breast cancer, colorectal cancer, rectal cancer, non-small cell lungcancer, non-Hodgkins lymphoma (NHL), renal cell cancer, prostate cancer,liver cancer, pancreatic cancer, soft-tissue sarcoma, Kaposi's sarcoma,carcinoid carcinoma, head and neck cancer, melanoma, ovarian cancer,mesothelioma, gliomas, glioblastomas, neuroblastomas, and multiplemyeloma. In some embodiments, the cancer is a solid tumor.

In some embodiments, the cancer to be treated is breast cancer, such asmetastatic breast cancer. In some embodiments, the cancer to be treatedis lung cancer, such as non-small cell lung cancer, including advancedstage non-small cell lung cancer. In some embodiments, the cancer to betreated is pancreatic cancer, such as early stage pancreatic cancer oradvanced or metastatic pancreatic cancer. In some embodiments, thecancer to be treated is melanoma, such as stage III or IV melanoma.

In some embodiments, the individual being treated for a proliferativedisease has been identified as having one or more of the conditionsdescribed herein. Identification of the conditions as described hereinby a skilled physician is routine in the art (e.g., via blood tests,X-rays, CT scans, endoscopy, biopsy, angiography, CT-angiography, etc.)and may also be suspected by the individual or others, for example, dueto tumor growth, hemorrhage, ulceration, pain, enlarged lymph nodes,cough, jaundice, swelling, weight loss, cachexia, sweating, anemia,paraneoplastic phenomena, thrombosis, etc. In some embodiments, theindividual has been identified as susceptible to one or more of theconditions as described herein. The susceptibility of an individual maybe based on any one or more of a number of risk factors and/ordiagnostic approaches appreciated by the skilled artisan, including, butnot limited to, genetic profiling, family history, medical history(e.g., appearance of related conditions), lifestyle or habits.

In some embodiments, the methods and/or compositions (such aspharmaceutical compositions) used herein reduce the severity of one ormore symptoms associated with proliferative disease (e.g., cancer) by atleast about any one of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%,or 100% compared to the corresponding symptom in the same individualprior to treatment or compared to the corresponding symptom in otherindividuals not receiving the methods and/or compositions (such aspharmaceutical compositions).

In some embodiments, the composition (such as a pharmaceuticalcomposition) (such as a pharmaceutical composition) described herein isused in combination with another administration modality or treatment.For example, in some embodiments, the composition (such as apharmaceutical composition) is used in combination with gemcitabine (forexample for treating pancreatic cancer). In some embodiments, thecomposition (such as a pharmaceutical composition) is used incombination with carboplatin (for example for treating lung cancer).

Dosing and Method of Administration

The amount of the pharmaceutical composition administered to anindividual (such as a human) may vary with the particular composition,the method of administration, and the particular type of recurrentcancer being treated. The amount should be sufficient to produce adesirable beneficial effect. For example, in some embodiments, theamount of the composition (such as a pharmaceutical composition) iseffective to result in an objective response (such as a partial responseor a complete response). In some embodiments, the amount ofpharmaceutical composition is sufficient to result in a completeresponse in the individual. In some embodiments, the amount of thecomposition (such as a pharmaceutical composition) is sufficient toresult in a partial response in the individual. In some embodiments, theamount of the composition (such as a pharmaceutical composition)administered alone is sufficient to produce an overall response rate ofmore than about any one of 40%, 50%, 60%, or 64% among a population ofindividuals treated with the composition (such as a pharmaceuticalcomposition). Responses of an individual to the treatment of the methodsdescribed herein can be determined, for example, based on RECIST orCA-125 level. For example, when CA-125 is used, a complete response canbe defined as a return to a normal range value of at least 28 days fromthe pretreatment value. A particle response can be defined as asustained over 50% reduction from the pretreatment value.

In some embodiments, the amount of pharmaceutical composition issufficient to prolong progress-free survival of the individual (forexample as measured by RECIST or CA-125 changes). In some embodiments,the amount of the pharmaceutical composition is sufficient to prolongoverall survival of the individual. In some embodiments, the amount ofthe composition (such as a pharmaceutical composition) is sufficient toproduce clinical benefit of more than about any one of 50%, 60%, 70%, or77% among a population of individuals treated with the composition (suchas a pharmaceutical composition).

In some embodiments, the amount of paclitaxel in the composition (suchas a pharmaceutical composition) is below the level that induces atoxicological effect (i.e., an effect above a clinically acceptablelevel of toxicity) or is at a level where a potential side effect can becontrolled or tolerated when the composition (such as a pharmaceuticalcomposition) is administered to the individual. In some embodiments, theamount of the composition (such as a pharmaceutical composition) isclose to a maximum tolerated dose (MTD) of the composition (such as apharmaceutical composition) following the same dosing regimen. In someembodiments, the amount of the composition (such as a pharmaceuticalcomposition) is more than about any one of 80%, 90%, 95%, or 98% of theMTD.

In some embodiments, the amount of paclitaxel and/or composition is anamount sufficient to decrease the size of a tumor, decrease the numberof cancer cells, or decrease the growth rate of a tumor by at leastabout any one of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or100% compared to the corresponding tumor size, number of cancer cells,or tumor growth rate in the same subject prior to treatment or comparedto the corresponding activity in other subjects not receiving thetreatment. Standard methods can be used to measure the magnitude of thiseffect, such as in vitro assays with purified enzyme, cell-based assays,animal models, or human testing.

In some embodiments, the amount of paclitaxel in the composition (suchas a pharmaceutical composition) is included in any one of the followingranges: about 0.5 mg to about 5 mg, about 5 mg to about 10 mg, about 10mg to about 15 mg, about 15 mg to about 20 mg, about 20 mg to about 25mg, about 20 mg to about 50 mg, about 25 mg to about 50 mg, about 50 mgto about 75 mg, about 50 mg to about 100 mg, about 75 mg to about 100mg, about 100 mg to about 125 mg, about 125 mg to about 150 mg, about150 mg to about 175 mg, about 175 mg to about 200 mg, about 200 mg toabout 225 mg, about 225 mg to about 250 mg, about 250 mg to about 300mg, about 300 mg to about 350 mg, about 350 mg to about 400 mg, about400 mg to about 450 mg, or about 450 mg to about 500 mg. In someembodiments, the amount of paclitaxel in the composition (such as apharmaceutical composition) (e.g., a unit dosage form) is in the rangeof about 5 mg to about 500 mg, such as about 30 mg to about 300 mg orabout 50 mg to about 200 mg. In some embodiments, the concentration ofthe paclitaxel in the composition (such as a pharmaceutical composition)is dilute (about 0.1 mg/ml) or concentrated (about 100 mg/ml), includingfor example any one of about 0.1 mg/ml to about 50 mg/ml, about 0.1mg/ml to about 20 mg/ml, about 1 mg/ml to about 10 mg/ml, about 2 mg/mlto about 8 mg/ml, about 4 mg/ml to about 6 mg/ml, or about 5 mg/ml. Insome embodiments, the concentration of the paclitaxel is at least aboutany one of 0.5 mg/ml, 1.3 mg/ml, 1.5 mg/ml, 2 mg/ml, 3 mg/ml, 4 mg/ml, 5mg/ml, 6 mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, 10 mg/ml, 15 mg/ml, 20 mg/ml,25 mg/ml, 30 mg/ml, 40 mg/ml, or 50 mg/ml.

Exemplary doses of paclitaxel in the pharmaceutical composition include,but are not limited to, about any one of 25 mg/m², 30 mg/m², 50 mg/m²,60 mg/m², 75 mg/m², 80 mg/m², 90 mg/m², 100 mg/m², 120 mg/m², 160 mg/m²,175 mg/m², 180 mg/m², 200 mg/m², 210 mg/m², 220 mg/m², 250 mg/m², 260mg/m², 300 mg/m², 350 mg/m², 400 mg/m², 500 mg/m², 540 mg/m², 750 mg/m²,1000 mg/m², or 1080 mg/m² of paclitaxel. In various embodiments, thecomposition (such as a pharmaceutical composition) includes less thanabout any one of 350 mg/m², 300 mg/m², 250 mg/m², 200 mg/m², 150 mg/m²,120 mg/m², 100 mg/m², 90 mg/m², 50 mg/m², or 30 mg/m² of paclitaxel. Insome embodiments, the amount of paclitaxel per administration is lessthan about any one of 25 mg/m², 22 mg/m², 20 mg/m², 18 mg/m², 15 mg/m²,14 mg/m², 13 mg/m², 12 mg/m², 11 mg/m², 10 mg/m², 9 mg/m², 8 mg/m², 7mg/m², 6 mg/m², 5 mg/m², 4 mg/m², 3 mg/m², 2 mg/m², or 1 mg/m². In someembodiments, the dose of paclitaxel in the composition (such as apharmaceutical composition) is included in any one of the followingranges: about 1 mg/m² to about 5 mg/m², about 5 mg/m² to about 10 mg/m²,about 10 mg/m² to about 25 mg/m², about 25 mg/m² to about 50 mg/m²,about 50 mg/m² to about 75 mg/m², about 75 mg/m² to about 100 mg/m²,about 100 mg/m² to about 125 mg/m², about 125 mg/m² to about 150 mg/m²,about 150 mg/m² to about 175 mg/m², about 175 mg/m² to about 200 mg/m²,about 200 mg/m² to about 225 mg/m², about 225 mg/m² to about 250 mg/m²,about 250 mg/m² to about 300 mg/m², about 300 mg/m² to about 350 mg/m²,or about 350 mg/m² to about 400 mg/m². Preferably, the dose ofpaclitaxel in the composition (such as a pharmaceutical composition) isabout 5 mg/m² to about 300 mg/m², such as about 100 mg/m² to about 150mg/m², about 120 mg/m², about 130 mg/m², or about 140 mg/m².

In some embodiments of any of the above aspects, the dose of paclitaxelin the composition (such as a pharmaceutical composition) includes atleast about any one of 1 mg/kg, 2.5 mg/kg, 3.5 mg/kg, 5 mg/kg, 6.5mg/kg, 7.5 mg/kg, 10 mg/kg, 15 mg/kg, or 20 mg/kg. In variousvariations, the dose of paclitaxel in the composition (such as apharmaceutical composition) includes less than about any one of 350mg/kg, 300 mg/kg, 250 mg/kg, 200 mg/kg, 150 mg/kg, 100 mg/kg, 50 mg/kg,25 mg/kg, 20 mg/kg, 10 mg/kg, 7.5 mg/kg, 6.5 mg/kg, 5 mg/kg, 3.5 mg/kg,2.5 mg/kg, 2 mg/kg, 1.5 mg/kg, or 1 mg/kg of paclitaxel. In someembodiments, the dose of paclitaxel in the composition (such as apharmaceutical composition) includes less than about any one of 500μg/kg, 350 μg/kg, 300 μg/kg, 250 μg/kg, 200 μg/kg, 150 μg/kg, 100 μg/kg,50 μg/kg, 25 μg/kg, 20 μg/kg, 10 μg/kg, 7.5 μg/kg, 6.5 μg/kg, 5 μg/kg,3.5 μg/kg, 2.5 μg/kg, 2 μg/kg, 1.5 μg/kg, 1 μg/kg, or 0.5 μg/kg ofpaclitaxel.

Exemplary dosing frequencies include, but are not limited to, any one ofweekly without break; weekly, three out of four weeks; once every threeweeks; once every two weeks; weekly, two out of three weeks. In someembodiments, the composition (such as a pharmaceutical composition) isadministered about once every 2 weeks, once every 3 weeks, once every 4weeks, once every 6 weeks, or once every 8 weeks. In some embodiments,the composition (such as a pharmaceutical composition) is administeredat least about any one of 1×, 2×, 3×, 4×, 5×, 6×, or 7× (i.e., daily) aweek. In some embodiments, the intervals between each administration areless than about any one of 6 months, 3 months, 1 month, 20 days, 15,days, 12 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days,3 days, 2 days, or 1 day. In some embodiments, the intervals betweeneach administration are more than about any one of 1 month, 2 months, 3months, 4 months, 5 months, 6 months, 8 months, or 12 months. In someembodiments, there is no break in the dosing schedule. In someembodiments, the interval between each administration is no more thanabout a week.

The administration of the composition (such as a pharmaceuticalcomposition) can be over an extended period of time, such as from abouta month up to about seven years. In some embodiments, the composition(such as a pharmaceutical composition) is administered over a period ofat least about any one of 2 months, 3 months, 4 months, 5 months, 6months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months,18 months, 24 months, 30 months, 36 months, 48 months, 60 months, 72months, or 84 months. In some embodiments, the composition (such as apharmaceutical composition) is administered over a period of at leastone month, wherein the interval between each administration is no morethan about a week, and wherein the dose of paclitaxel at eachadministration is about 0.25 mg/m² to about 75 mg/m², such as about 0.25mg/m² to about 25 mg/m² or about 25 mg/m² to about 50 mg/m².

In some embodiments, the dosage of paclitaxel in a pharmaceuticalcomposition can be in the range of 5 mg/m² to 400 mg/m² when given on a3 week schedule, or 5 mg/m² to 250 mg/m² when given on a weeklyschedule. For example, the amount of a paclitaxel is about 60 mg/m² toabout 300 mg/m² (e.g., about 260 mg/m²).

Other exemplary dosing schedules for the administration of thepharmaceutical composition include, but are not limited to, any one of100 mg/m², weekly, without break; 75 mg/m² weekly, 3 out of four weeks;100 mg/m², weekly, 3 out of 4 weeks; 125 mg/m², weekly, 3 out of 4weeks; 125 mg/m², weekly, 2 out of 3 weeks; 130 mg/m², weekly, withoutbreak; 175 mg/m², once every 2 weeks; 260 mg/m², once every 2 weeks; 260mg/m², once every 3 weeks; 180-300 mg/m², every three weeks; 60-175mg/m², weekly, without break; 20-150 mg/m² twice a week; and 150-250mg/m² twice a week. The dosing frequency of the composition (such as apharmaceutical composition) may be adjusted over the course of thetreatment based on the judgment of the administering physician.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is administered (e.g., intravenously) at 260 mg/m² everythree weeks. In some embodiments, the composition (such as apharmaceutical composition) is administered (e.g., intravenously) at 220mg/m², every three weeks. In some embodiments, the composition (such asa pharmaceutical composition) is administered (e.g., intravenously) at180 mg/m², every three weeks. In some embodiments, the composition (suchas a pharmaceutical composition) is administered (e.g., intravenously)at 200 mg/m², every three weeks. In some embodiments, the composition(such as a pharmaceutical composition) is administered (e.g.,intravenously) at 130 mg/m², every three weeks.

In some embodiments, the composition (such as a pharmaceuticalcomposition) is administered (e.g., intravenously) at 150 mg/m² on days1, 8, and 15 every 4 weeks. In some embodiments, the composition (suchas a pharmaceutical composition) is administered (e.g., intravenously)at 125 mg/m2 on days 1, 8, and 15 every 4 weeks. In some embodiments,the composition (such as a pharmaceutical composition) is administered(e.g., intravenously) at 100 mg/m² on days 1, 8, and 15 every 4 weeks.In some embodiments, the composition (such as a pharmaceuticalcomposition) is administered (e.g., intravenously) at 75 mg/m2 on days1, 8, and 15 every 4 weeks. In some embodiments, the composition (suchas a pharmaceutical composition) is administered (e.g., intravenously)at 50 mg/m² on days 1, 8, and 15 every 4 weeks.

The compositions (such as pharmaceutical compositions) described hereinallow infusion of the composition (such as a pharmaceutical composition)to an individual over an infusion time that is shorter than about 24hours. For example, in some embodiments, the composition (such as apharmaceutical composition) is administered over an infusion period ofless than about any one of 24 hours, 12 hours, 8 hours, 5 hours, 3hours, 2 hours, 1 hour, 30 minutes, 20 minutes, or 10 minutes. In someembodiments, the composition (such as a pharmaceutical composition) isadministered over an infusion period of about 30 minutes. In someembodiments, the composition (such as a pharmaceutical composition) isadministered over an infusion period between about 30 minutes to about40 minutes.

In some embodiments, the present application provides a method oftreating cancer in an individual by parenterally administering to theindividual (e.g., a human) an effective amount of a composition (such asa pharmaceutical composition) described herein. The present applicationalso provides a method of treating cancer in an individual byintravenous, intra-arterial, intramuscular, subcutaneous, inhalation,oral, intraperitoneal, nasally, or intra-tracheal administering to theindividual (e.g., a human) an effective amount of a paclitaxelpharmaceutical composition. In some embodiments, the route ofadministration is intraperitoneal. In some embodiments, the route ofadministration is intravenous, intra-arterial, intramuscular, orsubcutaneous. In various variations, about 5 mg to about 500 mg, such asabout 30 mg to about 300 mg or about 50 mg to about 500 mg, of thepaclitaxel is administered per dose. In some embodiments, the paclitaxelis the only pharmaceutically active agent for the treatment of cancerthat is contained in the composition (such as a pharmaceuticalcomposition).

Any of the compositions (such as pharmaceutical compositions) describedherein can be administered to an individual (such as human) via variousroutes, including, for example, intravenous, intra-arterial,intraperitoneal, intrapulmonary, oral, inhalation, intravesicular,intramuscular, intra-tracheal, subcutaneous, intraocular, intrathecal,transmucosal, transdermal, intratumoral, direct injection into the bloodvessel wall, intracranial, or intra-cavity. In some embodiments,sustained continuous release formulation of the composition (such as apharmaceutical composition) may be used. In one variation describedherein, nanoparticles (such as albumin nanoparticles) of the inventivecompositions (such as pharmaceutical compositions) can be administeredby any acceptable route including, but not limited to, orally,intramuscularly, transdermally, intravenously, through an inhaler orother air borne delivery systems and the like.

In some embodiments, the albumin-based paclitaxel-containingpharmaceutical compositions may be administered with a secondtherapeutic compound and/or a second therapy. The dosing frequency ofthe composition (such as a pharmaceutical composition) and the secondcompound may be adjusted over the course of the treatment based on thejudgment of the administering physician. In some embodiments, the firstand second therapies are administered simultaneously, sequentially, orconcurrently. When administered separately, the pharmaceuticalcomposition and the second compound can be administered at differentdosing frequency or intervals. For example, the composition (such as apharmaceutical composition) can be administered weekly, while a secondcompound can be administered more or less frequently. In someembodiments, sustained continuous release formulation ofpaclitaxel-containing nanoparticle and/or second compound may be used.Various formulations and devices for achieving sustained release areknown in the art. A combination of the administration configurationsdescribed herein can be used.

In some embodiments, the cancer is breast cancer (for example metastaticbreast cancer), and the composition (such as a pharmaceuticalcomposition) is administered at 260 mg/m² once every three weeks.

In some embodiments, the cancer is pancreatic cancer (for exampleadvanced pancreatic cancer, or adenocarcinoma of the pancreas), and thecomposition (such as a pharmaceutical composition) is administered at125 mg/m² weekly, three out of four weeks. In some embodiments, thecancer is pancreatic cancer (for example advanced pancreatic cancer),and the composition (such as a pharmaceutical composition) isadministered at 125 mg/m² weekly, three out of four weeks in combinationwith gemcitabine at 1000 mg/m².

In some embodiments, the cancer is lung cancer (for example non-smallcell lung cancer), and the composition (such as a pharmaceuticalcomposition) is administered at 100 mg/m² weekly. In some embodiments,the cancer is lung cancer (for example non-small cell lung cancer), andthe composition (such as a pharmaceutical composition) is administeredat 100 mg/m² weekly, such as on Days 1, 8, 15 of each three weeks cycle,in combination with carboplatin at AUC=6 mg·min/mL once every threeweeks, such as on Day 1 of each three weeks cycle.

Metronomic Therapy Regimens

The present invention also provides metronomic therapy regimens for anyof the methods of treatment and methods of administration describedherein. Exemplary metronomic therapy regimens and variations for the useof metronomic therapy regimens are discussed below and disclosed in U.S.application Ser. No. 11/359,286, filed Feb. 21, 2006, published as US2006/0263434A1 (such as those described in paragraphs [0138] to [0157]therein), which is hereby incorporated by reference in its entirety. Insome embodiments, the pharmaceutical composition is administered over aperiod of at least one month, wherein the interval between eachadministration is no more than about a week, and wherein the dose of thepaclitaxel at each administration is about 0.25% to about 25% of itsmaximum tolerated dose following a traditional dosing regimen. In someembodiments, the pharmaceutical composition is administered over aperiod of at least two months, wherein the interval between eachadministration is no more than about a week, and wherein the dose of thepaclitaxel at each administration is about 1% to about 20% of itsmaximum tolerated dose following a traditional dosing regimen. In someembodiments, the dose of paclitaxel per administration is less thanabout any one of 25%, 24%, 23%, 22%, 20%, 18%, 15%, 14%, 13%, 12%, 11%,10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the maximum tolerateddose. In some embodiments, any pharmaceutical composition isadministered at least about any one of 1×, 2×, 3×, 4×, 5×, 6×, or 7×(i.e., daily) a week. In some embodiments, the intervals between eachadministration are less than about any one of 6 months, 3 months, 1month, 20 days, 15, days, 12 days, 10 days, 9 days, 8 days, 7 days, 6days, 5 days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, theintervals between each administration are more than about any one of 1month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, or 12months. In some embodiments, the composition (such as a pharmaceuticalcomposition) is administered over a period of at least about any one of2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9months, 10 months, 11 months, 12 months, 18 months, 24 months, 30months, 36 months, 48 months, 60 months, 72 months, or 84 months.

EXEMPLARY EMBODIMENTS Embodiment 1

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin polymers among the albumin on thenanoparticles, wherein a percentage of albumin polymer among the albuminon the nanoparticles being about 15% to about 40% (such as about 15% toabout 20%, about 20% to about 24.5%, about 24.5% to about 30%, about 30%to about 35%, or about 35% to about 40%) is indicative of suitability ofthe pharmaceutical composition for medical use.

Embodiment 2

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe percentage of albumin monomers among the albumin on thenanoparticles, wherein a percentage of albumin monomers among thealbumin on the nanoparticles being about 40% to about 60% (such as about40% to about 55%, about 40% to about 54%, about 40% to about 53%, about40% to about 52%, about 40% to about 50%, about 40% to about 48%, orabout 40% to about 46%) is indicative of suitability of thepharmaceutical composition for medical use.

Embodiment 3

In some further embodiments of embodiment 1, the method furthercomprises determining the percentage of albumin monomers among thealbumin on the nanoparticles, wherein a percentage of albumin monomersamong the albumin on the nanoparticles being about 40% to about 60%(such as about 40% to about 55%, about 40% to about 54%, about 40% toabout 53%, about 40% to about 52%, about 40% to about 50%, about 40% toabout 48%, or about 40% to about 46%) is indicative of suitability ofthe pharmaceutical composition for medical use.

Embodiment 4

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight percentage of the albumin in the nanoparticles, wherein aweight percentage of the albumin in the nanoparticles being about 15% toabout 30% is indicative of suitability of the pharmaceutical compositionfor medical use.

Embodiment 5

In some further embodiments of any one of embodiments 1-3, the methodfurther comprises determining the weight percentage of the albumin inthe nanoparticles, wherein a weight percentage of the albumin in thenanoparticles being about 15% to about 30% is indicative of suitabilityof the pharmaceutical composition for medical use.

Embodiment 6

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe weight ratio of albumin to paclitaxel in the nanoparticles, whereinan albumin to paclitaxel ratio of about 1:2 to about 1:6 in thenanoparticles is indicative of suitability of the pharmaceuticalcomposition for medical use.

Embodiment 7

In some further embodiments of any one of embodiments 1-5, the methodfurther comprises determining the weight ratio of albumin to paclitaxelin the nanoparticles, wherein an albumin to paclitaxel ratio of about1:2 to about 1:6 in the nanoparticles is indicative of suitability ofthe pharmaceutical composition for medical use.

Embodiment 8

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe morphology of the nanoparticles under cryo-TEM, wherein an irregularshape of the nanoparticles is indicative of suitability of thepharmaceutical composition for medical use.

Embodiment 9

In some further embodiments of any one of embodiments 1-8, the methodfurther comprises determining the morphology of the nanoparticles undercryo-TEM, wherein an irregular shape of the nanoparticles is indicativeof suitability of the pharmaceutical composition for medical use.

Embodiment 10

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe thickness of the albumin coating of the nanoparticles undercryo-TEM, wherein a thickness of about 5-7 nm (such as about 6 nm) isindicative of suitability of the pharmaceutical composition for medicaluse.

Embodiment 11

In some further embodiments of any one of embodiments 1-9, the methodfurther comprises determining the thickness of the albumin coating ofthe nanoparticles under cryo-TEM, wherein a thickness of about 5-7 nm(such as about 6 nm) is indicative of suitability of the pharmaceuticalcomposition for medical use.

Embodiment 12

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe distribution of paclitaxel in a tumor tissue upon injection of thepharmaceutical composition directly into the tumor tissue; wherein anenhanced paclitaxel tumor distribution is indicative of suitability ofthe pharmaceutical composition for medical use.

Embodiment 13

In some further embodiments of any one of embodiments 1-11, the methodfurther comprises determining the distribution of paclitaxel in a tumortissue upon injection of the pharmaceutical composition directly intothe tumor; wherein an enhanced paclitaxel tumor distribution isindicative of suitability of the pharmaceutical composition for medicaluse.

Embodiment 14

In some embodiments, there is provided a method of assessing suitabilityof a pharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, the method comprising: determiningthe solubility, paclitaxel crystallinity, and a paclitaxel recoveryfollowing a 0.2 micron filtration of the pharmaceutical composition,wherein a solubility of about 50 μg/ml to about 80 μg/ml in a 5% humanalbumin solution, a non-crystalline state of the paclitaxel, and apaclitaxel recovery date of at least about 80% is indicative ofsuitability of the pharmaceutical composition for medical use.

Embodiment 15

In some further embodiments of any one of embodiments 1-13, the methodfurther comprises determining the solubility of the pharmaceuticalcomposition, wherein a solubility of about 50 μg/ml to about 80 μg/ml ina 5% human albumin solution is indicative of suitability of thepharmaceutical composition for medical use.

Embodiment 16

In some further embodiments of any one of embodiments 1-13 and 15, themethod further comprises determining the paclitaxel crystallinity of thepharmaceutical composition, wherein a non-crystalline state of thepaclitaxel is indicative of suitability of the pharmaceuticalcomposition for medical use.

Embodiment 17

In some further embodiments of any one of embodiments 1-13 and 15, themethod further comprises determining the paclitaxel recovery following a0.2 micron filtration of the pharmaceutical composition, wherein apaclitaxel recovery of at least about 80% is indicative of suitabilityof the pharmaceutical composition for medical use.

Embodiment 18

In some further embodiments of any one of embodiments 15-17, thedetermination of solubility, paclitaxel crystalline state, or paclitaxelrecovery is carried out after storage.

Embodiment 19

In some further embodiments of any one of embodiments 14 and 16-18, thepaclitaxel crystallinity is determined by X-ray diffraction, polarizedlight microscopy, or both.

Embodiment 20

In some further embodiments of any one of embodiments 1-19, the methodfurther comprises determining the binding affinity of albumin topaclitaxel in the pharmaceutical composition.

Embodiment 21

In some further embodiments of embodiment 20, the binding affinity isdetermined by equilibrium dialysis, FTIR, NMR, or a combination thereof.

Embodiment 22

In some further embodiments of any one of embodiments 1-21, the methodfurther comprises determining the surface-to-volume ratio of thenanoparticles in the pharmaceutical composition.

Embodiment 23

In some further embodiments of any one of embodiments 1-22, the methodfurther comprises determining the percentage of albumin dimers among thealbumin on the nanoparticles, wherein a percentage of about 15% to about30% of albumin dimers among the albumin on the nanoparticles isindicative of the pharmaceutical composition for medical use.

Embodiment 24

In some further embodiments of any one of embodiments 1-23, the methodfurther comprises determining the percentage of albumin oligomers amongthe albumin on the nanoparticles, wherein a percentage of about 7% toabout 15% of albumin oligomers among the albumin on the nanoparticles isindicative of the pharmaceutical composition for medical use.

Embodiment 25

In some further embodiments of any one of embodiments 1-24, the methodfurther comprises determining the percentage of albumin monomers,dimers, oligomers, or polymers among the total albumin in thepharmaceutical composition.

Embodiment 26

In some further embodiments of any one of embodiments 1-3, 5, 7, 9, 11,13, and 15-25, the percentage of albumin monomers, dimers, oligomers, orpolymers is carried out by size-exclusion chromatography.

Embodiment 27

In some further embodiments of any one of embodiments 1-26, the methodfurther comprises determining the particle size of the nanoparticles.

Embodiment 28

In some further embodiments of embodiment 27, the particle size of thenanoparticles is determined by dynamic light scattering.

Embodiment 29

In some further embodiments of any one of embodiments 1-28, the methodfurther comprises determining the polydispersity index of thenanoparticles in the pharmaceutical composition.

Embodiment 30

In some further embodiments of any one of embodiments 1-29, the methodfurther comprises determining the span of size distribution((Dv₉₀-Dv₁₀)/Dv₅₀) of the nanoparticles in the pharmaceuticalcomposition.

Embodiment 31

In some further embodiments of any one of embodiments 1-30, the methodfurther comprises determining the surface potential of thenanoparticles.

Embodiment 32

In some further embodiments of any one of embodiments 1-31, the methodfurther comprises determining the percentage of the paclitaxel in thenanoparticles among the total paclitaxel in the pharmaceuticalcomposition.

Embodiment 33

In some further embodiments of embodiment 32, the percentage of thepaclitaxel in the nanoparticles is determined by reversed-phase HPLC.

Embodiment 34

In some further embodiments of any one of embodiments 1-33, the methodfurther comprises determining the percentage of the albumin that is inthe non-nanoparticle portion among the total albumin in thepharmaceutical composition.

Embodiment 35

In some further embodiments of embodiment 34, the percentage of thealbumin is determined by size-exclusion chromatography.

Embodiment 36

In some further embodiments of any one of embodiments 1-35, the methodfurther comprises determining the stability of the pharmaceuticalcomposition.

Embodiment 37

In some further embodiments of embodiment 36, the stability isdetermined after storage.

Embodiment 38

In some further embodiments of any one of embodiments 1-11 and 14-37,the method further comprises determining tumor distribution ofpaclitaxel upon administration in vivo.

Embodiment 39

In some further embodiments of embodiment 38, the method comprisesdetermining tumor distribution of paclitaxel upon injection of thepharmaceutical composition directly into the tumor tissue.

Embodiment 40

In some further embodiments of any one of embodiments 1-39, the weightratio of the total albumin to the total paclitaxel in the pharmaceuticalcomposition is about 3:1 to about 7.9:1 or about 10:1 to about 17:1.

Embodiment 41

In some further embodiments of any one of embodiments 1-40, the albuminis human albumin.

Embodiment 42

In some further embodiments of any one of embodiments 1-41, the averageparticle size of the nanoparticles is less than about 200 nm.

Embodiment 43

In some embodiments, there is provided a method of validating acommercial batch of a pharmaceutical composition for medical use in ahuman individual, wherein the pharmaceutical composition comprisesnanoparticles comprising paclitaxel coated with albumin and anon-nanoparticle portion comprising albumin and paclitaxel, and whereinthe method comprises 1) obtaining a sample from the commercial batch,and 2) assessing suitability of the sample for medical use according toany one of embodiments 1-42.

Embodiment 44

In some embodiments, there is provided a commercial batch of apharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises nanoparticlescomprising paclitaxel coated with albumin and a non-nanoparticle portioncomprising albumin and paclitaxel, and wherein the commercial batch isvalidated by assessment of suitability for medical use according to anyone of embodiments 1-42.

EXAMPLES

The examples below are intended to be purely exemplary of the inventionand should therefore not be considered to limit the invention in anyway. The following examples and detailed description are offered by wayof illustration and not by way of limitation.

Example 1. Determination of the Distribution of Paclitaxel within TumorTissue

This example demonstrates the measurement of the distribution ofpaclitaxel in tumor tissue. Distribution of paclitaxel activity was usedto monitor drug penetration and tumor cell uptake at defined radialdistances extending from a site of microinjection for three formulationsof paclitaxel.

To generate xenografts, athymic Nude-Foxnlnu mice (Jackson Laboratories)were injected subcutaneously with 2.5×10⁶ MIA PaCa-2, A2058 melanoma, orH2122 non-small cell lung cancer (NSCLC) cells in a 1:1 ratio with BDBiosciences Matrigel Matrix.

Three formulations of paclitaxel were prepared for this study:paclitaxel (PTX) solubilized in DMSO (PTX:DMSO), paclitaxel solubilizedin oil-based solvent Cremophor EL (PTX:CrEL), and a nanoparticleformulation of paclitaxel and albumin sold under the trademark ABRAXANE®(ABX). The concentration of paclitaxel in each formulation wasdetermined by liquid chromatography mass spectrometry (LC-MS).

Microinjections were performed using the CIVO™ arrayed microinjectiondevice (Presage Biosciences, Seattle, Wash.) by inserting the devicetranscutaneously into flank tumors of anesthetized mice. A minimum of 3tumors per time point were used with 2-3 replicate injection sites performulation in each tumor. An average drug volume of 3 μL was deliveredvia an extrusion method over an injection column length of 6 mm. Anequal amount of paclitaxel in each formulation (12 μg) was administeredper injection. Inactivated near infrared dye, VivoTag680-S (50 μg/mL),was co-injected with each drug to mark the injection site.

Tumors were analyzed 24, 48, or 72 hours post-drug microinjection formitotic arrest by immunofluorescent staining of phospho-histone H3(pHH3). At these post-injection time points, animals were euthanized.Tumors were harvested and resected, fixed in 10% buffered formalin for48 hours, and scanned on a Xenogen IVIS in the near infrared spectrum(excitation 680 nm, emission 720 nm) to confirm the location of eachinjection site. Each tumor was cut into 2 mm thick cross sectionsperpendicular to the plane of injection to enable a three-dimensionalassessment of the entire injection column.

Following IVIS imaging, tumors were processed for standard paraffinembedding and histological analysis. 4 micron sections, cut from each 2mm cross section, were stained with an anti-pHH3 antibody and an AlexaFluor 488 secondary antibody to assess drug-induced tumor responses(mitotic arrest) using custom software (CIVOanalyzer™; PresageBiosciences, Seattle, Wash.). Tissue sections were stained with4′,6-diamindino-2-phenylindole (DAPI) to visualize nuclei. Mean fractionvalues of pHH3 positive cells were plotted with standard error bars, asa function of radial distance for each formulation and time point. Toassess the statistical significance of the differences between any pairof formulations, a linear mixed model approach was used. In the model,the response to the PTX:CrEL formulation was assumed to be a randomeffect and the differential response due to the nab-paclitaxel soldunder the trademark ABRAXANE® or PTX:DMSO was assumed to be a fixedeffect. A p-value of less than 0.05, adjusted for multiple comparisons,indicates statistically significant differences. Data are expressed asmean plus/minus standard error.

Immunofluorescent staining of pHH3 was used as a pharmacodynamicsindicator of paclitaxel activity to monitor drug penetration and tumorcell uptake for MIA PaCa-2 xenografts at defined radial distancesextending from the site of injection. FIGS. 1A-1I show representativeimaging of pancreatic MIA PaCa-2 at 24, 48, and 72 hours followingmicroinjection with ABX, PTX:DMSO, or PTX:CrEL. At all three time points(24, 48, and 72 hours), the area of response and the total fraction ofpHH3 positive cells at a specific radial distance following drugmicroinjection were significantly greater for the nab-paclitaxel soldunder the trademark ABRAXANE® compared with either PTX:DMSO or PTX:CrEL(p<0.01)(FIGS. 2A-2C).

Microinjections of paclitaxel formulations were also tested on twoadditional tumor xenografts, namely, A2058 melanoma and H2122 NSCLCxenografts. Results for microinjection of A2058 xenografts showed thatABX induced a larger increase in both the area of response and totalfraction of cells arrested in mitosis at 24 hours post-injection whencompared to PTX:CrEL (FIG. 3A)(n=5 tumors; p<0.001). Results formicroinjection of H2122 xenografts showed that ABX induced a largerincrease in both the area of response and total fraction of cellsarrested in mitosis at 24 hours post-injection when compared to PTX:DMSOmicroinjection of (FIG. 3B)(n=3 tumors; p<0.001).

Example 2. Determination of the Distribution of Paclitaxel within TumorTissue for Increasing Concentrations of Paclitaxel

This example demonstrates the measurement of the distribution ofpaclitaxel in tumor tissue. Distribution of paclitaxel activity was usedto monitor drug penetration and tumor cell uptake at defined radialdistances extending from a site of microinjection for three formulationsof paclitaxel at three paclitaxel concentrations.

MIA PaCa-2 xenografts were generated as discussed in Example 1.

Three paclitaxel formulations were prepared for this study: paclitaxel(PTX) solubilized in DMSO (PTX:DMSO), paclitaxel solubilized inoil-based solvent Cremophor EL (PTX:CrEL), and a nanoparticleformulation of paclitaxel and albumin sold under the trademark ABRAXANE®(ABX). The concentration of paclitaxel in each formulation wasdetermined by liquid chromatography mass spectrometry (LC-MS).

Microinjections were performed using the CIVO™ arrayed microinjectiondevice (Presage Biosciences, Seattle, Wash.) by inserting the devicetranscutaneously into flank tumors of anesthetized mice. A minimum of 3tumors per time point were used with 2-3 replicated injection sites performulation in each tumor. An average drug volume of 3 μL was deliveredvia an extrusion method over an injection column length of 6 mm.Paclitaxel, as measured in PTX:DMSO, PTX:CrEL, and ABX, was administeredat three concentrations: 1.6 mg/mL, 2.5 mg/mL, or 4.75 mg/mL.Inactivated near infrared dye, VivoTag680-S (50 μg/mL), was co-injectedwith each drug to mark the injection site.

Tumors were analyzed at 24 hours post-drug microinjection for mitoticarrest by immunofluorescent staining of phospho-histone H3 (pHH3). Tumortissue samples were prepared and analyzed as discussed in Example 1.

Immunofluorescent staining of pHH3 was used as a pharmacodynamicsindicator of paclitaxel activity to monitor drug penetration and tumorcell uptake at defined radial distances extending from the site ofinjection for 3 concentrations of paclitaxel. At 24 hours, the area ofresponse and the total fraction of pHH3 positive cells at a specificradial distance were greater for the three concentrations ofmicroinjected nab-paclitaxel sold under the trademark ABRAXANE® comparedwith respective concentrations of either PTX:DMSO and PTX:CrEL (FIGS.4A-4C).

Example 3. Ultracentrifugation of Nanoparticles in the Composition

This example demonstrates a method for separating nanoparticles in thecomposition (such as a pharmaceutical composition) from anon-nanoparticle portion of a composition (such as paclitaxel andalbumin). Ultracentrifugation was performed at speeds and durations thatallow the sedimentation of the nanoparticles without significantsedimentation of any albumin and/or paclitaxel not associated with thenanoparticles.

A pharmaceutical composition comprising nanoparticles comprisingpaclitaxel coated with albumin and a non-nanoparticle portion comprisingalbumin and paclitaxel was obtained. The composition was reconstitutedwith 0.9% sodium chloride to yield a 5 mg/mL suspension as measured bypaclitaxel. 4.0 mL aliquots of reconstituted suspension were transferredto Beckman Coulter quick seal polyallomer bell-top tubes and submittedto ultracentrifugation at 50,000 rpm for 41 minutes at 25° C. in a Type00 Ti rotor. After ultracentrifugation, the tubes were removed withoutdisturbing the pelleted nanoparticles. A micropipette was used to remove3.0 mL of supernatant from each tube. Each supernatant was transferredto a separate test tube and saved for further analysis. The remainingsupernatant was poured out of the tube containing the pelletednanoparticles. The pellet was then gently washed with 2 mL of water. Thewater was poured out of the tube and the pellet was again gently washedwith 2 mL of water.

Example 4. Determining the Concentration of Paclitaxel and Albumin in aNon-Nanoparticle Portion of a Composition

This example demonstrates a method for measuring the concentration ofpaclitaxel and albumin from a non-nanoparticle portion of a composition(such as a pharmaceutical composition).

As demonstrated in Example 3, the non-nanoparticle portion of apharmaceutical composition was separated from the nanoparticle portionof the pharmaceutical composition by ultracentrifugation. To determinethe concentration of albumin in the non-nanoparticle portion of thepharmaceutical composition, 1.0 mL of the supernatant obtained followingultracentrifugation was transferred to a 10-mL volumetric flask. Thesupernatant was then diluted to 10-mL with 0.9% sodium chloride. Thisdilute solution was then subjected to HPLC analysis as discussed below.

Briefly, human albumin standards were created with a concentration of0.4 mg/mL in 0.9% sodium chloride. An HPLC system equipped with avariable wavelength UV/VIS detector and data acquisition system was setup with a TosoHaas TSK Guard Column SWxL (6.0 mm×40 mm, 7 μm particlesize) and a TosoHaas Analytical Column G3000 SWxL (7.8 mm×300 mm, 5 μmparticle size) kept at ambient temperature. Separate 10 μL injections ofthe albumin standards or dilute supernatant samples obtained fromultracentrifugation of the pharmaceutical composition were analyzed onthe HPLC system at 228 nm using a 60-minute chromatography cycle with aflow rate of 1.0 mL/min using 100 mM K₂HPO₄ (pH 7.0) mobile phase. Aftereach chromatographic cycle, the entire line of the HPLC system,including pump and columns, were washed with a 0.05% sodium azidesolution. The concentration of albumin in the non-nanoparticle portionof the pharmaceutical composition was calculated using the informationobtained from the chromatograms of the human albumin standards and thesamples.

To determine the concentration of paclitaxel in the non-nanoparticleportion of the pharmaceutical composition, 1.0 mL of the supernatantobtained following ultracentrifugation was transferred to a 25-mLvolumetric flask. The supernatant was then diluted to 25.0 mL withpurified water, sonicated for 5 minutes, and then allowed to cool toroom temperature. 5.0 mL of this solution was then transferred into a10-mL volumetric flask, diluted to 10.0 mL with a 50:50 solution ofacetonitrile and water, sonicated for 5 minutes, and then allowed tocool to room temperature. This solution was then subjected to HPLCanalysis as discussed below.

Briefly, paclitaxel standards were created with a concentration of 1.2mg/mL in acetonitrile. 2.0 mL of the 1.2 mg/mL paclitaxel solution wastransferred to a 100-mL volumetric flask and diluted to volume withacetonitrile to obtain a solution of 24 μg/mL paclitaxel. 2.0 mL of the24 μg/mL paclitaxel solution was transferred to a 25-mL volumetric flaskand diluted to volume with acetonitrile to obtain a solution of 1.9μg/mL paclitaxel. This standard solution was stored at 4° C. until use.Additionally, a system sensitivity solution of paclitaxel was prepared.2.0 mL of the 1.9 μg/mL paclitaxel solution was transferred to a 50-mLvolumetric flask and diluted to volume with acetonitrile to obtain asolution of 0.08 μg/mL paclitaxel. An HPLC system equipped with a UVabsorbance detector and data acquisition system was set up with aPhenomenex, Curosil PFP guard column (4.6 mm×30 mm, 5 μm particle size)and a Phenomenex, Curosil PFP analytical column (4.6 mm×250 mm, 5 μmparticle size). The autosampler was maintained at 4° C. Separate 10 μLinjections of the paclitaxel system sensitivity solution, paclitaxelstandard or dilute supernatant samples obtained from ultracentrifugationof the pharmaceutical composition were analyzed on the HPLC system at228 nm using a 10-minute chromatography cycle with a flow rate of 1.0mL/min using a 70:30 acetonitrile:water mobile phase. To check thesystem suitability for sample injection, the 1.9 μg/mL paclitaxelstandard was injected and analyzed, followed by an injection andanalysis of 100% acetonitrile, and then an injection and analysis of thesystem sensitivity solution of paclitaxel. The height of theinterference peak in the acetonitrile was confirmed to be not more thanone-fourth of the paclitaxel peak height in the analysis of the systemsensitivity solution of paclitaxel. Additionally, it was confirmed thatthe signal-to-noise ratio of the paclitaxel peak in the systemsensitivity solution of paclitaxel was not less than 10. Theconcentration of paclitaxel in the non-nanoparticle portion of thepharmaceutical composition was calculated using the information obtainedfrom the chromatograms of the paclitaxel standard and the samples.

Example 5. Determining the Concentration of Paclitaxel and Albumin in aNanoparticle Portion of a Composition

This example demonstrates a method for measuring the concentration ofpaclitaxel and albumin from a nanoparticle portion of a composition(such as a pharmaceutical composition).

As demonstrated in Example 3, the nanoparticle portion of apharmaceutical composition was separated from the non-nanoparticleportion of the pharmaceutical composition by ultracentrifugation. Todetermine the concentration of albumin in the nanoparticle portion ofthe pharmaceutical composition, 3.0 mL of ethanol (200 absolute proof)was added to the pellet and sonicated until the pellet was fullydispersed. After the pellet was dispersed, the solution was transferredwith a glass pipette to another centrifuge tube. The original tube wasrinsed with 2 mL ethanol and transferred to the other centrifuge tube.The sample was then centrifuged at 10,000 rpm for 20 minutes at 25° C.After centrifugation, the ethanol was removed with a long glass pipette.The pellet was then dried in a desiccator under vacuum for about 1-2hours. 3.0 mL of 0.9% sodium chloride was added to the dried pellet andthen sonicated to disperse the pellet into a homogeneous mixture. Themixture was then transferred into a 10-mL volumetric flask, using 5.0 mLof 0.9% sodium chloride to wash and transfer the mixture, and thendiluted to volume using 0.9% sodium chloride. The solution was sonicateduntil dissolved into a homogeneous solution and then allowed to cool toroom temperature. This solution was then subjected to HPLC analysis asdiscussed in Example 4.

As demonstrated in Example 3, the nanoparticle portion of apharmaceutical composition was separated from the non-nanoparticleportion of the pharmaceutical composition by ultracentrifugation. Todetermine the concentration of paclitaxel in the nanoparticle portion ofthe pharmaceutical composition, 3.0 mL of a 50:50 acetonitrile:watersolution was added to the pellet and then sonicated until the pelletdispersed into a homogeneous mixture. The mixture was transferred into a250-mL volumetric flask and additional 50:50 acetonitrile:water solutionwas used to wash the tube that contained the pellet. The solution wasthen diluted to volume using the 50:50 acetonitrile:water solution,sonicated until the pellet was completely dissolved, and then allowed tocool to room temperature. 2.0 mL of this solution was transferred into a100-mL volumetric flask and diluted to volume with the 50:50acetonitrile:water solution. This solution was then subjected to HPLCanalysis as discussed in Example 4.

Example 6. Determining the Percentage of Albumin Monomers, Dimers,Oligomers, and Polymers Among the Albumin on the Nanoparticles in theComposition

This example demonstrates the measurement of albumin monomers, dimers,oligomers, and polymers among the total albumin either in thenanoparticle portion of the pharmaceutical composition or in thenon-nanoparticle portion of the pharmaceutical composition.

HPLC analysis of albumin in a nanoparticle portion of a composition andalbumin in a non-nanoparticle portion of a composition was performed asdiscussed herein. Chromatograms were generated as discussed above. Forexample, A chromatogram from the method was generated as shown in FIG.5. The RRT, as compared to monomeric albumin, for the separated albuminspecies was determined and used to classify peaks in the chromatographsas monomeric, dimeric, oligomeric, or polymeric albumin. The percentageof albumin monomers, dimers, oligomers, and polymers in the nanoparticleportion of the composition was calculated by comparing the integratedpeak areas of each albumin species to the total integrated peak area ofthe albumin on the nanoparticles.

Example 7. Determining the Weight Percentage of Albumin and Paclitaxelin the Nanoparticles in the Composition

This example demonstrates the measurement of weight percentage ofalbumin on the nanoparticles and paclitaxel in the nanoparticles in thepharmaceutical composition.

A pharmaceutical composition comprising nanoparticles comprisingpaclitaxel coated with albumin and a non-nanoparticle portion comprisingalbumin and paclitaxel was obtained. If necessary, the composition wasreconstituted. The nanoparticles in the composition were then isolatedaway from the non-nanoparticle portion of the composition byultracentrifugation as discussed above.

Isolated nanoparticles were diluted and the amount of albumin andpaclitaxel in the nanoparticles was measured. The amount of albumin onthe nanoparticle is measured as detailed herein. The amount ofpaclitaxel was measured by a RP-HPLC method as detailed herein.Paclitaxel was detected at 228 nm.

The weight percentage of albumin on the nanoparticles was determinedfrom the amount of albumin on the nanoparticles and the total weight ofthe nanoparticles (e.g., the amount of albumin and the amount ofpaclitaxel). The weight ratio of albumin on the nanoparticles topaclitaxel in the nanoparticles was determined from the amount ofalbumin on the nanoparticles and the amount of paclitaxel in thenanoparticles.

Example 8. Determining the Morphology and Thickness of the Nanoparticlesin the Composition Using Cryo-TEM

This example demonstrates a cryo-TEM technique for determining themorphology and thickness of the nanoparticles in the pharmaceuticalcomposition.

A pharmaceutical composition comprising nanoparticles comprisingpaclitaxel coated with albumin and a non-nanoparticle portion comprisingalbumin and paclitaxel was obtained. If necessary, the composition wasreconstituted. Optionally, the nanoparticles in the composition are thenisolated away from the non-nanoparticle portion of the composition byultracentrifugation as discussed above.

Nanoparticles were prepared for cryo-TEM imaging. In short, thereconstituted nanoparticles were rapidly cooled to cryogenictemperatures to form a vitreous form of the reconstituted compositionwhich was then analyzed. Briefly, the nanoparticles of the compositionwere analyzed in their native structure at magnifications ranging from6,500× to 110,000× at different areas in the sample. The morphology ofthe nanoparticles, such as the irregularity of shape, rugosity, andsurface-to-volume ratio, was assessed from the cryo-TEM images. Thethickness of the albumin coating on the nanoparticles was also measuredfrom the cryo-TEM image.

Example 9. Determining the Solubility, Paclitaxel Crystallinity, andPaclitaxel Recovery of the Composition Following 0.2 Micron Filtrationof the Composition

This example demonstrates the measurement of nanoparticle solubility,paclitaxel crystallinity, and paclitaxel recovery of the pharmaceuticalcomposition following 0.2 micron filtration.

A pharmaceutical composition comprising nanoparticles comprisingpaclitaxel coated with albumin and a non-nanoparticle portion comprisingalbumin and paclitaxel was obtained. If necessary, the composition wasreconstituted. The composition was assessed for nanoparticle solubility,paclitaxel crystallinity, and paclitaxel recovery of the pharmaceuticalcomposition following 0.2 micron filtration immediately afterreconstitution and after an accelerated aging process. To age thecomposition, the reconstituted composition was stored for 24 hours at40° C. Compositions were filtered by passing the reconstitutedcomposition through a 0.2 micron filter.

Solubility of nanoparticles of the composition was determined byperforming dynamic light scattering measurements on a series ofconcentrations of the composition after reconstitution. The proportionof intact particles to free paclitaxel was a function of the solubilityof the particles. Thus, as measured by this method, the solubility wasdetermined as the concentration below which particles were no longerdetectable by dynamic light scattering.

Paclitaxel crystallinity of the nanoparticles of the composition wasdetermined by performing an X-ray diffraction method and a polarizedlight microscopy method after reconstitution. Optionally, X-raydiffraction measurements were made on isolated nanoparticles.Nanoparticles in the composition were optionally isolated away from thenon-nanoparticle portion of the composition by ultracentrifugation asdiscussed herein. The isolated nanoparticles were then optionally driedby lyophilization. Non-crystalline paclitaxel in the nanoparticles willexhibit broad scattering halos, indicative of an amorphous material(e.g., non-crystalline). Crystalline paclitaxel in the nanoparticleswill exhibit numerous well-defined scattering peaks. Polarized lightmicroscopy measurements were performed on a suspension of thecomposition. To determine the crystalline state of the nanoparticles ofthe composition, a birefringence test was performed with an opticalmicroscope using polarized light.

Paclitaxel recovery following 0.2 micron filtration of the compositionwas determined by performing RP-HPLC. Following 0.2 micron filtration,nanoparticles in the composition were optionally isolated away from thenon-nanoparticle portion of the composition by ultracentrifugation asdiscussed herein. The amount of paclitaxel in the nanoparticlesrecovered after the 0.2 micron filtration was measured by a RP-HPLCmethod as discussed herein. Paclitaxel was detected at 228 nm. Thedegree of paclitaxel recovery was assessed by comparing the amount ofpaclitaxel in the composition following 0.2 micron filtration to theamount of paclitaxel in the composition prior to 0.2 micron filtration.

Example 10. Validating a Composition for Medical Use

This example demonstrates the validation of a composition for medicaluse by assessing particle morphology, particle size, surface potential,paclitaxel crystallinity, fraction of free and bound paclitaxel oralbumin, nature of the bond between paclitaxel and albumin, dissolutionprofile, and oligomeric status of albumin.

A pharmaceutical composition comprising nanoparticles comprisingpaclitaxel coated with albumin and a non-nanoparticle portion comprisingalbumin and paclitaxel was obtained. If necessary, the composition wasreconstituted. The composition was assessed for sameness, compared tonab-paclitaxel sold under the trademark ABRAXANE®, with respect toparticle morphology, particle size (both Dv₅₀ and span or polydispersityindex), surface potential, paclitaxel crystallinity, fraction of freeand bound paclitaxel or albumin, nature of the bond between paclitaxeland albumin, dissolution profile, and oligomeric status of albumin.

To assess for sameness with respect to particle morphology ofnanoparticles in the pharmaceutical composition, the morphology of thenanoparticles was assessed.

To assess for sameness with respect to particle size and span orpolydispersity index of the nanoparticles in the pharmaceuticalcomposition, the reconstituted composition was subjected to dynamiclight scattering measurements. The particle size was measured as themean particle size (D₅₀) of the nanoparticles in the composition. Thespan or polydispersity index was measured as the span of the sizedistribution, (D₉₀−D₁₀)/D₅₀.

To assess for sameness with respect to the surface potential ofnanoparticles in the pharmaceutical composition, the zeta potential ofthe nanoparticles was determined. The zeta potential was measured usingtechniques such as microelectrophoresis, electrophoretic lightscattering, dynamic electrophoretic mobility, or tunable resistive pulsesensing (TRPS).

To assess for sameness with respect to paclitaxel crystallinity ofnanoparticles in the pharmaceutical composition, the crystallinity ofpaclitaxel in the nanoparticles of the composition was assessed asdiscussed above.

To assess for sameness with respect to the fraction of free and boundpaclitaxel or albumin in the pharmaceutical composition, thenanoparticle portion of the composition was separated from thenon-nanoparticle portion of the composition. Nanoparticles in thecomposition are isolated away from the non-nanoparticle portion of thecomposition by ultracentrifugation as discussed herein. Isolatednanoparticles are diluted and the amount of albumin or paclitaxel in thenanoparticles was measured. The amount of albumin on the nanoparticlewas measured by a HPLC size-exclusion chromatography method. Albumin wasdetected at 228 nm. The amount of paclitaxel was measured by a RP-HPLCmethod. Paclitaxel was detected at 228 nm. The non-nanoparticle portionof the composition is isolated and if the amount of albumin wasdetermined in the nanoparticles, then the amount of albumin wasdetermined in the non-nanoparticle portion of the composition. Theamount of albumin in the non-nanoparticle portion of the composition wasmeasured by a HPLC size-exclusion chromatography method. Albumin wasdetected at 228 nm. If the amount of paclitaxel was determined in thenanoparticles, then the amount of paclitaxel was determined in thenon-nanoparticle portion of the composition. The amount of paclitaxelwas measured by a RP-HPLC method. Paclitaxel was detected at 228 nm. Thefraction of free (non-nanoparticle portion) and bound paclitaxel(nanoparticle portion) or albumin was calculated from the measuredvalues.

To assess for sameness with respect to the nature of the bond betweenpaclitaxel and albumin, the nature of the bond between albumin andpaclitaxel was assessed.

To assess for sameness with respect to the dissolution profile, thereconstituted composition was subjected to an in vitro release kineticsassay. The reconstituted composition was diluted in a 0.9% salinesolution and dynamic light scattering was used to monitor particle sizeover 60 minutes for a range of concentrations of the composition, or thereconstituted composition was diluted in a 5% human albumin solution anddynamic light scattering was used to monitor scattering and particlesize over 60 minutes for a range of concentrations of the composition,or the reconstituted composition was diluted in water and a UV-Visspectrophotometer was used to monitor absorbance over 60 minutes for arange of concentrations of the composition.

To assess for sameness with respect to the oligomeric status of albumin,albumins in the starting material and in the final composition wereassessed for oligomeric status. Isolated albumin in the startingmaterial and final composition were analyzed by a HPLC size-exclusionchromatography method. Albumin was detected at 228 nm. A chromatogram ofthe HPLC-size exclusion chromatography method was generated as shown inFIG. 5. The RRT, as compared to monomeric albumin, for the separatedalbumin species was determined and used to classify peaks in thechromatographs as monomeric, dimer, oligomeric, or polymeric albumin.

The composition was suitable for medical use if the particle morphology,particle size, surface potential, paclitaxel crystallinity, fraction offree and bound paclitaxel or albumin, nature of the bond betweenpaclitaxel and albumin, dissolution profile, and oligomeric status ofalbumin were found to exhibit sameness with respect to results obtainedfrom analysis of nab-paclitaxel sold under the trademark ABRAXANE®.

Example 11. Validating a Composition for Medical Use

This example demonstrates the validation of a composition for medicaluse, both before and after storage, by assessing particle morphology,particle size, surface potential, paclitaxel crystallinity, fraction offree and bound paclitaxel or albumin, nature of the bond betweenpaclitaxel and albumin, dissolution profile, oligomeric status ofalbumin, oligomeric status of albumin on the nanoparticle, and therecovery of paclitaxel following 0.2 micron filtration.

A pharmaceutical composition comprising nanoparticles comprisingpaclitaxel coated with albumin and a non-nanoparticle portion comprisingalbumin and paclitaxel was obtained. If necessary, the composition wasreconstituted. The composition was assessed for sameness, in view ofnab-paclitaxel sold under the trademark ABRAXANE®, with respect toparticle morphology, particle size, surface potential, paclitaxelcrystallinity, fraction of free and bound paclitaxel or albumin, natureof the bond between paclitaxel and albumin, dissolution profile,oligomeric status of albumin, oligomeric status of albumin on thenanoparticle, and the recovery of paclitaxel following 0.2 micronfiltration. To test the composition following storage conditions, thecomposition was stored for 24 hours at 40° C. to simulate an acceleratedaging process.

To assess for sameness with respect to particle morphology ofnanoparticles in the pharmaceutical composition, the nanoparticles inthe composition were assessed as discussed herein. The morphology of thenanoparticles, such as the irregularity of shape, was assessed from thecryo-TEM images.

To assess for sameness with respect to particle size and span orpolydispersity index of the nanoparticles in the pharmaceuticalcomposition, the reconstituted composition was subjected to dynamiclight scattering measurements. The particle size was measured as thevolume-weighted mean particle size (Dv₅₀) of the nanoparticles in thecomposition. The span or polydispersity index was measured as the spanof the volume-weighted size distribution, (Dv₉₀−Dv₁₀)/Dv₅₀.

To assess for sameness with respect to the surface potential ofnanoparticles in the pharmaceutical composition, the zeta potential ofthe nanoparticles was determined. The zeta potential was measured usingtechniques such as microelectrophoresis, electrophoretic lightscattering, dynamic electrophoretic mobility, or tunable resistive pulsesensing (TRPS).

To assess for sameness with respect to paclitaxel crystallinity ofnanoparticles in the pharmaceutical composition, paclitaxelcrystallinity of the nanoparticles of the composition was determined byperforming an X-ray diffraction method and a polarized light microscopymethod as detailed herein.

To assess for sameness with respect to the fraction of free (i.e., notassociated with the nanoparticles) and bound (i.e., associated with thenanoparticles) paclitaxel and albumin in the pharmaceutical composition,the composition was assessed as detailed herein.

To assess for sameness with respect to the nature of the bond betweenpaclitaxel and albumin, the composition was assessed via equilibriumdialysis test and FTIR and NMR analysis. The nature of the bond betweenpaclitaxel and albumin was assessed using paclitaxel and processedalbumin. The binding affinity between paclitaxel and processed albuminin the composition was measured via an equilibrium dialysis testingapparatus and further by FTIR and NMR analysis.

To assess for sameness with respect to the dissolution profile, thereconstituted composition was subject to an in vitro release kineticsassay. The reconstituted composition was diluted in a 0.9% salinesolution and dynamic light scattering is used to monitor particle sizeover 60 minutes for a range of concentrations of the composition, or thereconstituted composition was diluted in a 5% human albumin solution anddynamic light scattering was used to monitor scattering and particlesize over 60 minutes for a range of concentrations of the composition,or the reconstituted composition was diluted in water and a UV-Visspectrophotometer was used to monitor absorbance over 60 minutes for arange of concentrations of the composition.

To assess for sameness with respect to the oligomeric status of albumin,albumins in the starting material and in the final composition areisolated away from the other components of the composition and theiroligomeric status was assessed. Isolated albumins in the startingproduct of final composition are analyzed by a HPLC size-exclusionchromatography method as detailed herein.

To assess for sameness with respect to the oligomeric status of albuminon the nanoparticle, the nanoparticles in the composition were isolatedaway from the non-nanoparticle portion of the composition byultracentrifugation as discussed herein. Isolated nanoparticles arediluted and then analyzed by a HPLC size-exclusion chromatography methodas discussed herein.

To assess for sameness with respect to the recovery of paclitaxelfollowing 0.2 micron filtration, the reconstituted composition wasfiltered with a 0.2 micron filter. Following 0.2 micron filtration,nanoparticles in the composition are isolated away from thenon-nanoparticle portion of the composition by ultracentrifugation asdiscussed herein. The amount of paclitaxel in the recoverednanoparticles from the 0.2 micron filtered composition was measured by aRP-HPLC method as discussed herein. Paclitaxel was detected at 228 nm.The degree of paclitaxel recovery was assessed by comparing the amountof paclitaxel in the composition following 0.2 micron filtration to theamount of paclitaxel in the composition prior to 0.2 micron filtration.

The composition was suitable for medical use if the particle morphology,particle size, surface potential, paclitaxel crystallinity, fraction offree and bound paclitaxel or albumin, nature of the bond betweenpaclitaxel and albumin, dissolution profile, oligomeric status ofalbumin, status of albumin on the nanoparticle, and the recovery ofpaclitaxel following 0.2 micron filtration were found to exhibitsameness with respect to results obtained from analysis ofnab-paclitaxel sold under the trademark ABRAXANE®.

Example 12. Determining In Vitro Dissolution Kinetics of thePharmaceutical Composition Using a UV-Vis Spectrophotometer

This example demonstrates the measurement of in vitro release kineticsof the pharmaceutical composition after reconstitution, as determined bya UV-Vis spectrophotometer.

The pharmaceutical composition was reconstituted with a 0.9% sodiumchloride solution to produce a 5 mg/ml stock solution of thepharmaceutical composition, as measured by the concentration ofpaclitaxel. For example, starting from a vial of the lyophilizedpharmaceutical composition containing 100 mg, as measured by the amountof paclitaxel, 20 ml of 0.9% sodium chloride was slowly injected intothe vial over a minimum of 1 minute using a sterile syringe. The flow of0.9% sodium chloride was directed onto the inside wall of the vial.Subsequently, the lyophilized pharmaceutical composition was allowed torest for 5 minutes and then the vial was gently swirled or slowlyinverted for at least 2 minutes until complete dissolution of thepharmaceutical composition occurred. The lyophilized pharmaceuticalcomposition was reconstituted in a manner to avoid the formation offoam. The spectrophotometer was set up as shown in FIG. 6. A 295 nmlongwave pass filter was placed between the UV light source and thecuvette. A 10-mm quartz cuvette was placed into an Agilent Cary 8454UV-Vis spectrophotometer pre-equilibrated to 20° C. while stirring at2000 rpm. An appropriate volume of water was transferred to the cuvette.A magnetic stirrer bar was added to the bottom of cuvette. Thespectrophotometer was equilibrated over an hour (or longer) until the340 nm signal was stabilized. An appropriate volume of the stockreconstituted pharmaceutical composition suspension was then added tothe cuvette to achieve a target paclitaxel concentration of 100 μg/ml.The suspension was immediately mixed and the cuvette was capped.

During the preparation of the 100 μg/ml pharmaceutical composition, aseries of consecutive intensity measurement was performed. All kineticmeasurements were performed at 20° C. with the following settings: pathlength: 1 cm; wavelength range: 190-1100 nm; integration time: 0.5seconds; interval: 1 nm; deuterium lamp (UV): On; run time: 3600seconds; cycle time: 0.5 seconds; wavelength: 340 nm; and stirrer speed:2000 rpm.

As shown in FIG. 7, the majority of particles in the reconstitutedpharmaceutical composition suspension diluted to concentrations abovethe paclitaxel solubility in the release media dissolved anddisintegrated rapidly, first over 30 seconds, followed by a long slowtransient release over 3600 seconds or longer. The rate ofdisintegration and dissolution, and the dissolution profile shapes wereconcentration dependent.

Example 13. Methods of Making a Nanoparticle Composition

This example demonstrates methods of making an albumin/paclitaxelnanoparticle composition (such as a pharmaceutical composition) andvariants thereof.

All variants were manufactured using laboratory scale/bench-topequipment.

Variant 5 (V5) was prepared using the following procedure.

5% human albumin (HA) solution and paclitaxel solvent solutioncontaining approximately 200 mg/mL paclitaxel in 90:10 v:vchloroform:ethanol mixture were prepared. 18.4 mL of the 5% HA solutionwas transferred to a beaker and mixed using a Silverson high-shearmixer. 1.6 mL of the paclitaxel solvent solution was slowly (drop-wise)added to the HA solution and mixed for approximately 5 minutes at5000-6000 rpm to create a crude emulsion. The crude emulsion washigh-pressure homogenized at pressure of approximately 18-20 kpsi usingAvestin EmulsiFlex-C5 emulsifier for approximately 12 passes to create afine emulsion. The fine emulsion was transferred to a 2 L round bottomflask and the solvents were removed using BUCHI rotary evaporator with awater bath temperature set at 40° C. The evaporation was performed usingthe parameters in Table 1. The evaporation continued until the initialvolume was reduced by approximately 45-80%. The whole process wasrepeated once to generate enough material for in-process (IP) andfinished product (FP) testing. The post-evaporated (PE) suspension wascombined, mixed and assayed for paclitaxel and HA. Based on the assayvalues, the PE suspension was diluted with MilliQ water and 20-25% HAsolution to paclitaxel concentration of 7 mg/mL and HA concentration of56 mg/mL. The diluted suspension was filtered through a series of 1.2μm, 0.8 μm, 0.45 μm and 0.2 μm syringe filters with Supor PES membrane.1-3 mL aliquots of the filtered suspension were filled in 10 mL, 20 mmglass vials and lyophilized in VirTis Genesis EL25 lyophilizer (SPIndustries, Gardiner, N.Y.) using the cycle in Table 2. Afterlyophilization the vials were stoppered under nitrogen, crimped andstored at −20° C. for future testing.

A total of three lots of Variant 5 were manufactured.

TABLE 1 Evaporation cycle. Step # Pressure set point Hold time afterpressure is achieved 1 70 mm Hg 1 min 2 60 mm Hg 1 min 3 50 mm Hg 1 min4 40 mm Hg 1 min 5 30 mm Hg 1 min 6 25 mm Hg As needed

TABLE 2 Lyophilization cycle. Step Temp. (° C.) Time (min) Vac (mTorr)Type Loading 1 −55 N/A N/A Hold Freezing 1 −55 240 N/A Hold Drying 1 −5510 350 Hold 2 −15 200 350 Ramp 3 −15 10 350 Hold 4 25 400 350 Ramp 5 25840 350 Hold 6 30 50 350 Ramp 7 30 480 350 Hold

Variant 1 (V1) was prepared using the procedure for Variant 5 with thefollowing modification. The organic solvent mixture used to prepare the200 mg/mL paclitaxel solvent solution contained 50% by volume ethanoland 50% by volume chloroform.

A total of three lots of Variant 1 were manufactured.

Variant 2 (V2) was prepared using the procedure for Variant 5 with thefollowing modification. The concentration of the human albumin solutionused to prepare the crude emulsion was 10 mg/mL.

A total of three lots of Variant 2 were manufactured.

Variant 3 (V3) was prepared using the procedure for Variant 5 with thefollowing modification. The high-pressure homogenization of the crudeemulsion was performed at pressure of approximately 5 kpsi.

A total of three lots of Variant 3 were manufactured.

Variant 4 (V4) was prepared using the procedure for Variant 5 with thefollowing modifications. After the high-pressure homogenization the fineemulsion was transferred to a 500 mL round bottom flask and the solventswere removed using BUCHI rotary evaporator with a water bath temperatureset at 30° C. The evaporation was performed using the parameters inTable 3.

TABLE 3 Evaporation cycle for variant 4. Step # Pressure set point Holdtime after pressure is achieved 1 70 mm Hg 1 min 2 60 mm Hg 1 min 3 50mm Hg 1 min 4 From 40 mm Hg to 1 min 25 mm Hg, every 1 mm Hg 5 15 mm HgAs needed

A total of three lots of Variant 4 were manufactured.

Example 14. Assessment of Compositions

This example reports results from the assessment of variousalbumin/paclitaxel nanoparticle compositions.

The compositions were assessed using the methods described herein.Variants are designated as discussed herein (e.g., FP refers to finishedproduct; IP refers to in-process product). ABRAXANE® is the proprietaryalbumin/paclitaxel nanoparticle product of Celgene/Abraxis. PaclitaxelNAB, Albupax, and PacliALL are purported copies of the proprietaryalbumin/paclitaxel nanoparticle product of Celgene/Abraxis sold underthe trademark ABRAXANE® made by different companies.

TABLE 4 D_(V4,3) (nm) measured immediately after reconstitution. SampleName N (number) Mean Margin of error Min Max ABRAXANE ® 22 153.8 2.8144.1 164.4 Paclitax NAB 3 113.7 12.7 108.5 118.7 Albupax 5 140.0 12.0129.6 151.6 PacliALL 5 426.3 864.0 221.7 827.9 V5 FP 3 159.5 19.8 153.4168.5 V1 FP 3 177.6 74.4 158.8 212.1 V2 FP 3 179.5 27.1 167.0 186.9 V3FP 3 178.9 17.8 171.5 185.8 V4 FP 3 147.2 9.8 142.7 150.0

TABLE 5 Z average (nm) measured immediately after reconstitution. SampleName N (number) Mean Margin of error Min Max ABRAXANE ® 22 145.9 1.9138.5 153.0 Paclitax NAB 3 118.7 8.5 115.2 122.0 Albupax 5 138.9 1.7137.2 140.5 PacliALL 5 178.6 13.6 165.8 190.0 V5 FP 3 149.7 12.2 146.5155.4 V1 FP 3 164.8 55.7 149.3 190.5 V2 FP 3 161.4 12.9 155.6 165.6 V3FP 3 162.6 6.8 159.9 165.4 V4 FP 3 142.9 6.7 139.9 145.1

TABLE 6 Polydispersity index (PDI) measured immediately afterreconstitution. Sample Name N (number) Mean Margin of error Min MaxABRAXANE ® 22 0.119 0.006 0.097 0.145 Paclitax NAB 3 0.099 0.039 0.0870.117 Albupax 5 0.117 0.011 0.109 0.132 PacliALL 5 0.166 0.060 0.1180.241 V5 FP 3 0.127 0.027 0.118 0.139 V1 FP 3 0.092 0.079 0.071 0.129 V2FP 3 0.128 0.059 0.109 0.155 V3 FP 3 0.111 0.041 0.093 0.126 V4 FP 30.098 0.024 0.088 0.107

TABLE 7 D_(V5) (nm) measured immediately after reconstitution. SampleName N (number) Mean Margin of error Min Max ABRAXANE ® 22 75.0 2.3 68.288.0 Paclitax NAB 3 62.1 12.0 59.3 67.7 Albupax 5 73.4 19.8 55.2 91.8PacliALL 5 86.7 6.1 82.1 93.6 V5 FP 3 71.7 10.1 67.7 75.8 V1 FP 3 91.153.4 71.4 114.0 V2 FP 3 80.7 18.1 72.3 85.2 V3 FP 3 85.6 3.7 84.2 87.2V4 FP 3 76.2 8.9 73.3 80.2

TABLE 8 D_(V50) (nm) measured immediately after reconstitution. SampleName N (number) Mean Margin of error Min Max ABRAXANE ® 22 137.9 2.6 129148 Paclitax NAB 3 102.8 13.8 98.3 109 Albupax 5 130.6 15.3 117 146PacliALL 5 192.0 25.6 168 214 V5 FP 3 140.7 22.3 135 151 V1 FP 3 164.083.1 139 202 V2 FP 3 160.7 17.4 154 168 V3 FP 3 163.3 10.0 159 167 V4 FP3 134.3 10.0 130 138

TABLE 9 D_(V95) (nm) measured immediately after reconstitution. SampleName N (number) Mean Margin of error Min Max ABRAXANE ® 22  288.4 10.1237 341 Paclitax NAB 3 207.3 23.6 198 217 Albupax 5 243.2  8.5 235 251PacliALL 5 1263.0  2393.5  333 4710  V5 FP 3 314.0 33.4 299 325 V1 FP 3315.3 93.2 277 352 V2 FP 3 344.0 123.0  294 393 V3 FP 3 328.0 66.1 300353 V4 FP 3 264.3 22.5 256 274

TABLE 10 (D_(V90) − D_(V10))/D_(V50) measured immediately afterreconstitution. Sample Name N (number) Mean Margin of error Min MaxABRAXANE ® 22  1.21 0.06 0.82 1.53 Paclitax NAB 3 1.08 0.25 0.98 1.18Albupax 5 1.05 0.30 0.79 1.36 PacliALL 5 4.35 8.40 1.11 16.46  V5 FP 31.35 0.20 1.30 1.45 V1 FP 3 1.10 0.60 0.94 1.38 V2 FP 3 1.30 0.63 1.081.58 V3 FP 3 1.17 0.23 1.07 1.26 V4 FP 3 1.10 0.12 1.05 1.15

TABLE 11 Paclitaxel in solution phase (non-nanoparticle portion) as afraction of total paclitaxel, expressed as a percentage, immediatelyafter reconstitution. Sample Name N (number) Mean Margin of error MinMax ABRAXANE ® 65  1.71 0.027 1.46 2.14 Paclitax NAB 1 2.64 — 2.64 2.64Albupax 1 1.42 — 1.42 1.42 PacliALL 4 2.24 0.706 1.65 2.64 V5 FP 1 1.55— 1.55 1.55 V1 FP 1 1.51 — 1.51 1.51 V2 FP 1 1.63 — 1.63 1.63 V3 FP 11.55 — 1.55 1.55 V4 FP 1 1.59 — 1.59 1.59 V5 IP 1 0.85 — 0.85 0.85 V1 IP1 0.93 — 0.93 0.93 V2 IP 1 0.69 — 0.69 0.69 V3 IP 1 0.71 — 0.71 0.71 V4IP 1 1.23 — 1.23 1.23

TABLE 12 Paclitaxel in particles (nanoparticle portion) as a fraction oftotal paclitaxel, expressed as a percentage, immediately afterreconstitution. Sample Name N (number) Mean Margin of error Min MaxABRAXANE ® 65  98.290 0.025 97.860 98.540 Paclitax NAB 1 97.351 — 97.35197.351 Albupax 1 98.575 — 98.575 98.575 PacliALL 4 97.760 0.705 97.36098.350 V5 FP 1 98.441 — 98.441 98.441 V1 FP 1 98.482 — 98.482 98.482 V2FP 1 98.365 — 98.365 98.365 V3 FP 1 98.442 — 98.442 98.442 V4 FP 198.409 — 98.409 98.409 V5 IP 1 99.1  — 99.1  99.1  V1 IP 1 99.1  — 99.1 99.1  V2 IP 1 99.3  — 99.3  99.3  V3 IP 1 99.3  — 99.3  99.3  V4 IP 198.8  — 98.8  98.8 

TABLE 13 Albumin in solution phase (non-nanoparticle portion) as afraction of total albumin, expressed as a percentage, immediately afterreconstitution. Sample Name N (number) Mean Margin of error Min MaxABRAXANE ® 66  96.150 0.135 95.330 97.760 Paclitax NAB 1 98.701 — 98.70198.701 Albupax 1 96.648 — 96.648 96.648 PacliALL 4 98.450 0.175 98.36098.600 V5 FP 1 97.094 — 97.094 97.094 V1 FP 1 98.033 — 98.033 98.033 V2FP 1 97.961 — 97.961 97.961 V3 FP 1 96.846 — 96.846 96.846 V4 FP 196.033 — 96.033 96.033 V5 IP 1 94.1  — 94.1  94.1  V1 IP 1 96.9  — 96.9 96.9  V2 IP 1 71.0  — 71.0  71.0  V3 IP 1 90.8  — 90.8  90.8  V4 IP 195.6  — 95.6  95.6 

TABLE 14 Albumin in particles (nanoparticle portion) as a fraction oftotal albumin, expressed as a percentage, immediately afterreconstitution. Margin of Sample Name N (number) Mean error Min MaxABRAXANE ® 66  3.846 0.134 2.240 4.670 Paclitax NAB 1 1.299 — 1.2991.299 Albupax 1 3.352 — 3.352 3.352 PacliALL 4 1.555 0.173 1.400 1.640V5 FP 1 2.906 — 2.906 2.906 V1 FP 1 1.967 — 1.967 1.967 V2 FP 1 2.039 —2.039 2.039 V3 FP 1 3.154 — 3.154 3.154 V4 FP 1 3.967 — 3.967 3.967 V5IP 1 5.9  — 5.9 5.9 V1 IP 1 3.1  — 3.1 3.1 V2 IP 1 29.0   — 29.0 29.0 V3IP 1 9.2  — 9.2 9.2 V4 IP 1 4.4  — 4.4 4.4

TABLE 15 Concentration of free (in solution phase) paclitaxel (μg/ml) inthe composition immediately after reconstitution. Sample Name N (number)Mean Margin of error Min Max ABRAXANE ® 65   80.6  1.2  68.4  94.2Paclitax NAB 1 125.0 — 125.0 125.0 Albupax 1  71.5 —  71.5  71.5PacliALL 4 101.1 30.6  73.9 116.1 V5 FP 1  75.0 —  75.0  75.0 V1 FP 1 70.4 —  70.4  70.4 V2 FP 1  71.6 —  71.6  71.6 V3 FP 1  70.6 —  70.6 70.6 V4 FP 1  77.5 —  77.5  77.5 V5 IP 1 130.4 — 130.4 130.4 V1 IP 1115.8 — 115.8 115.8 V2 IP 1  71.2 —  71.2  71.2 V3 IP 1 102.0 — 102.0102.0 V4 IP 1 125.0 — 125.0 125.0

TABLE 16 Concentration of bound (in particles/nanoparticle portion)paclitaxel (μg/ml) in the composition immediately after reconstitution.Margin of Sample Name N (number) Mean error Min Max ABRAXANE ® 66  4624  45 4156  5147  Paclitax NAB 1 4594  — 4594  4594  Albupax 1 4945  —4945  4945  PacliALL 4 4405  176 4281  4547  V5 FP 1 4737  — 4737  4737 V1 FP 1 4568  — 4568  4568  V2 FP 1 4307  — 4307  4307  V3 FP 1 4461  —4461  4461  V4 FP 1 4795  — 4795  4795  V5 IP 1 15058.0 — 15058.015058.0 V1 IP 1 12282.4 — 12282.4 12282.4 V2 IP 1 10164.6 — 10164.610164.6 V3 IP 1 14152.6 — 14152.6 14152.6 V4 IP 1  9967.2 —  9967.2 9967.2

TABLE 17 Concentration of free (in solution phase) albumin (mg/ml) inthe composition immediately after reconstitution. Sample Name N (number)Mean Margin of error Min Max ABRAXANE ® 66  39.0 0.485 34.9 46.4Paclitax NAB 1 60.8 — 60.8 60.8 Albupax 1 34.4 — 34.4 34.4 PacliALL 442.7 1.005 41.8 43.3 V5 FP 1 40.1 — 40.1 40.1 V1 FP 1 36.6 — 36.6 36.6V2 FP 1 44.5 — 44.5 44.5 V3 FP 1 36.9 — 36.9 36.9 V4 FP 1 37.4 — 37.437.4 V5 IP 1 51.4 — 51.4 51.4 V1 IP 1 47.8 — 47.8 47.8 V2 IP 1  7.3 — 7.3  7.3 V3 IP 1 52.9 — 52.9 52.9 V4 IP 1 51.5 — 51.5 51.5

TABLE 18 Concentration of bound (in particles/nanoparticle portion)albumin (mg/ml) in the composition immediately after reconstitution.Sample Name N (number) Mean Margin of error Min Max ABRAXANE ® 66  1.5550.052 0.800 1.900 Paclitax NAB 1 0.800 — 0.800 0.800 Albupax 1 1.193 —1.193 1.193 PacliALL 4 0.676 0.087 0.595 0.715 V5 FP 1 1.199 — 1.1991.199 V1 FP 1 0.735 — 0.735 0.735 V2 FP 1 0.926 — 0.926 0.926 V3 FP 11.201 — 1.201 1.201 V4 FP 1 1.544 — 1.544 1.544 V5 IP 1 3.234 — 3.2343.234 V1 IP 1 1.520 — 1.520 1.520 V2 IP 1 2.996 — 2.996 2.996 V3 IP 15.376 — 5.376 5.376 V4 IP 1 2.382 — 2.382 2.382

TABLE 19 Albumin as a percentage of the nanoparticle mass immediatelyafter reconstitution. Sample Name N (number) Mean Margin of error MinMax ABRAXANE ® 30  24.0 1.4 13.6 29.0 Paclitax NAB 1 14.8 — 14.8 14.8Albupax 1 19.4 — 19.4 19.4 PacliALL 4 13.3 1.7 11.9 14.3 V5 FP 1 20.2 —20.2 20.2 V1 FP 1 13.9 — 13.9 13.9 V2 FP 1 17.7 — 17.7 17.7 V3 FP 1 21.2— 21.2 21.2 V4 FP 1 24.4 — 24.4 24.4 V5 IP 1 17.7 — 17.7 17.7 V1 IP 111.0 — 11.0 11.0 V2 IP 1 22.8 — 22.8 22.8 V3 IP 1 27.5 — 27.5 27.5 V4 IP1 19.3 — 19.3 19.3

A comparison of the albumin as a percentage of the nanoparticle mass(Table 19) is illustrated in the bar graph of FIG. 8. Amongst all thecompositions, ABRAXANE® and Paclitax NAB are most stable.

TABLE 20 Paclitaxel as a percentage of the nanoparticle mass immediatelyafter reconstitution. Sample Name N (number) Mean Margin of error MinMax ABRAXANE ® 30  76.1 1.435 71.0 86.4 Paclitax NAB 1 85.2 — 85.2 85.2Albupax 1 80.6 — 80.6 80.6 PacliALL 4 86.7 1.695 85.7 88.1 V5 FP 1 79.8— 79.8 79.8 V1 FP 1 86.1 — 86.1 86.1 V2 FP 1 82.3 — 82.3 82.3 V3 FP 178.8 — 78.8 78.8 V4 FP 1 75.6 — 75.6 75.6 V5 IP 1 82   — 82   82   V1 IP1 89   — 89   89   V2 IP 1 77   — 77   77   V3 IP 1 72   — 72   72   V4IP 1 81   — 81   81  

TABLE 21 Percentage of albumin in the form of monomers on thenanoparticles. Sample Name N (number) Mean Margin of error Min MaxABRAXANE ® 66  47.1  2.5 20.3 66.9 Paclitax NAB 1 44.8 — 44.8 44.8Albupax 1 55.6 — 55.6 55.6 PacliALL 4 54.1 20.2 40.7 70.9 V5 FP 1 68.5 —68.5 68.5 V1 FP 1 72.8 — 72.8 72.8 V2 FP 1 61.9 — 61.9 61.9 V3 FP 1 56.3— 56.3 56.3 V4 FP 1 63.5 — 63.5 63.5 V5 IP 1 66.1 — 66.1 66.1 V1 IP 178.1 — 78.1 78.1 V2 IP 1 51.9 — 51.9 51.9 V3 IP 1 55.7 — 55.7 55.7 V4 IP1 69.9 — 69.9 69.9

A comparison of the percentage of albumin on the nanoparticles in theform of monomers (Table 21) is illustrated in the bar graph of FIG. 9.Amongst all the compositions, ABRAXANE® and Paclitax NAB are moststable.

TABLE 22 Percentage of albumin in the form of dimers on thenanoparticles. Sample Name N (number) Mean Margin of error Min MaxABRAXANE ® 66  17.8 1.1  6.7 25.7 Paclitax NAB 1 19.4 — 19.4 19.4Albupax 1 16.5 — 16.5 16.5 PacliALL 4 21.9 6.0 16.6 25.1 V5 FP 1 11.0 —11.0 11.0 V1 FP 1 11.3 — 11.3 11.3 V2 FP 1 12.9 — 12.9 12.9 V3 FP 1  8.0—  8.0  8.0 V4 FP 1  9.8 —  9.8  9.8 V5 IP 1 13.5 — 13.5 13.5 V1 IP 110.9 — 10.9 10.9 V2 IP 1 21.0 — 21.0 21.0 V3 IP 1 11.6 — 11.6 11.6 V4 IP1 10.8 — 10.8 10.8

A comparison of the percentage of albumin on the nanoparticles in theform of dimers (Table 22) is illustrated in the bar graph of FIG. 10.Amongst all the compositions, the nab-paclitaxel sold under thetrademark ABRAXANE® and Paclitax NAB are most stable.

TABLE 23 Percentage of albumin in the form of oligomers on thenanoparticles. Sample Name N (number) Mean Margin of error Min MaxABRAXANE ® 66  7.9 0.9 3.1 16.5  Paclitax NAB 1 12.3  — 12.3  12.3 Albupax 1 9.1 — 9.1 9.1 PacliALL 4 13.4  6.8 8.3 17.2  V5 FP 1 3.2 — 3.23.2 V1 FP 1 3.3 — 3.3 3.3 V2 FP 1 4.0 — 4.0 4.0 V3 FP 1 2.6 — 2.6 2.6 V4FP 1 2.7 — 2.7 2.7 V5 IP 1 5.4 — 5.4 5.4 V1 IP 1 3.0 — 3.0 3.0 V2 IP 110.4  — 10.4  10.4  V3 IP 1 2.7 — 2.7 2.7 V4 IP 1 3.4 — 3.4 3.4

A comparison of the percentage of albumin on the nanoparticles in theform of oligomers (Table 23) is illustrated in the bar graph of FIG. 11.Amongst all the compositions, the nab-paclitaxel sold under thetrademark ABRAXANE® and Paclitax NAB are most stable.

TABLE 24 Percentage of albumin in the form of polymers on thenanoparticles. N Margin Sample Name (number) Mean of error Min MaxABRAXANE ® 66 27.2 2.5 10.7 69.0 Paclitax NAB 1 23.5 — 23.5 23.5 Albupax1 18.8 — 18.8 18.8 PacliALL 4 10.6 9.6 4.3 18.5 V5 FP 1 17.3 — 17.3 17.3V1 FP 1 12.7 — 12.7 12.7 V2 FP 1 21.1 — 21.1 21.1 V3 FP 1 33.2 — 33.233.2 V4 FP 1 24.0 — 24.0 24.0 V5 IP 1 15.0 — 15.0 15.0 V1 IP 1 8.0 — 8.08.0 V2 IP 1 16.8 — 16.8 16.8 V3 IP 1 30.1 — 30.1 30.1 V4 IP 1 15.9 —15.9 15.9

A comparison of the percentage of albumin on the nanoparticles in theform of polymers (Table 24) is illustrated in the bar graph of FIG. 12.Amongst all the compositions, the nab-paclitaxel sold under thetrademark ABRAXANE® and Paclitax NAB are most stable.

Using the measurements of albumin in the form of monomers (M), dimers(D), oligomers (O), and polymers (P) on the nanoparticles, compositionattributes were calculated as reported in Table 25.

TABLE 25 Summary of attributes calculated for albumin forms on thenanoparticles in the compositions. (P + O)/ (P + O)/ Sample Name D/M O/MP/M M (M − D) M + D M − D M + O M + P M − P D + O D + P O + P P/D O/DP/O ABRAXANE ® 37.7 16.8 57.7 74.6 119.7 64.9 29.4 55.1 74.3 19.9 25.745.0 35.1 153.3 44.7 343.0 Paclitax NAB 43.3 27.5 52.5 79.9 140.9 64.225.4 57.1 68.3 21.3 31.7 42.9 35.8 121.1 63.4 191.1 Albupax 29.8 16.433.8 50.2 71.5 72.1 39.0 64.7 74.3 36.8 25.7 35.3 27.9 113.5 55.1 205.9PacliALL 40.5 24.8 19.6 44.4 74.6 76.0 32.2 67.5 64.8 43.5 35.3 32.524.0 48.5 61.2 79.3 V5 FP 16.0 4.7 25.3 30.0 35.7 79.5 57.6 71.7 85.851.2 14.2 28.3 20.5 157.8 29.5 535.6 V1 FP 15.5 4.5 17.4 21.9 25.9 84.061.5 76.0 85.5 60.1 14.5 24.0 16.0 112.7 29.0 389.3 V2 FP 20.9 6.5 34.140.6 51.4 74.8 49.0 65.9 83.1 40.8 17.0 34.1 25.2 163.5 31.1 525.9 V3 FP14.2 4.5 58.9 63.4 73.9 64.3 48.3 58.9 89.5 23.2 10.5 41.1 35.7 416.132.1 1295.3 V4 FP 15.4 4.2 37.8 42.0 49.7 73.3 53.7 66.2 87.5 39.5 12.533.8 26.7 244.5 27.4 891.8 V5 IP 20.4 8.1 22.7 30.8 38.7 79.6 52.7 71.581.2 51.1 18.9 28.5 20.4 111.3 39.7 280.0 V1 IP 14.0 3.8 10.2 14.0 16.389.0 67.2 81.1 86.1 70.2 13.9 18.9 11.0 72.7 27.4 265.0 V2 IP 40.4 20.032.3 52.3 87.6 72.9 31.0 62.3 68.7 35.2 31.3 37.7 27.1 80.0 49.5 161.4V3 IP 20.8 4.8 54.0 58.8 74.3 67.2 44.1 58.4 85.8 25.6 14.3 41.6 32.8260.1 23.3 1117.8 V4 IP 15.5 4.9 22.7 27.6 32.7 80.7 59.0 73.3 85.8 54.014.2 26.7 19.3 146.7 31.5 466.0

A comparison of the percentage of albumin on the nanoparticles in theform of monomers (M) and dimers (D) (Table 25) is illustrated in the bargraph of FIG. 13. Amongst all the compositions, the nab-paclitaxel soldunder the trademark ABRAXANE® and Paclitax NAB are most stable.

A comparison of the percentage of albumin on the nanoparticles in theform of monomers (M) minus the percentage of albumin on thenanoparticles in the form of dimers (D) (Table 25) is illustrated in thebar graph of FIG. 14. Amongst all the compositions, the nab-paclitaxelsold under the trademark ABRAXANE® and Paclitax NAB are most stable.

A comparison of the percentage of albumin on the nanoparticles in theform of monomers (M) and oligomers (O) (Table 25) is illustrated in thebar graph of FIG. 15. Amongst all the compositions, the nab-paclitaxelsold under the trademark ABRAXANE® and Paclitax NAB are most stable.

A comparison of the percentage of albumin on the nanoparticles in theform of monomers (M) and polymers (P) (Table 25) is illustrated in thebar graph of FIG. 16. Amongst all the compositions, the nab-paclitaxelsold under the trademark ABRAXANE® and Paclitax NAB are most stable.

A comparison of the percentage of albumin on the nanoparticles in theform of monomers (M) minus the percentage of albumin on thenanoparticles in the form of polymers (P) (Table 25) is illustrated inthe bar graph of FIG. 17. Amongst all the compositions, thenab-paclitaxel sold under the trademark ABRAXANE® and Paclitax NAB aremost stable.

A comparison of the percentage of albumin on the nanoparticles in theform of dimers (D) and oligomers (O) (Table 25) is illustrated in thebar graph of FIG. 18. Amongst all the compositions, the nab-paclitaxelsold under the trademark ABRAXANE® and Paclitax NAB are most stable.

A comparison of the percentage of albumin on the nanoparticles in theform of dimers (D) and polymers (P) (Table 25) is illustrated in the bargraph of FIG. 19. Amongst all the compositions, the nab-paclitaxel soldunder the trademark ABRAXANE® and Paclitax NAB are most stable.

A comparison of the percentage of albumin on the nanoparticles in theform of oligomers (O) and polymers (P) (Table 25) is illustrated in thebar graph of FIG. 20. Amongst all the compositions, the nab-paclitaxelsold under the trademark ABRAXANE® and Paclitax NAB are most stable.

A comparison of the ratio (as reported as a percentage) of thepercentage of albumin on the nanoparticles in the form of dimers (D)divided by the percentage of albumin on the nanoparticles in the form ofmonomers (M) (Table 25) is illustrated in the bar graph of FIG. 21.Amongst all the compositions, the nab-paclitaxel sold under thetrademark ABRAXANE® and Paclitax NAB are most stable.

A comparison of the ratio (as reported as a percentage) of thepercentage of albumin on the nanoparticles in the form of oligomers (O)divided by the percentage of albumin on the nanoparticles in the form ofmonomers (M) (Table 25) is illustrated in the bar graph of FIG. 22.Amongst all the compositions, the nab-paclitaxel sold under thetrademark ABRAXANE® and Paclitax NAB are most stable.

A comparison of the ratio (as reported as a percentage) of thepercentage of albumin on the nanoparticles in the form of polymers (P)divided by the percentage of albumin on the nanoparticles in the form ofmonomers (M) (Table 25) is illustrated in the bar graph of FIG. 23.Amongst all the compositions, the nab-paclitaxel sold under thetrademark ABRAXANE® and Paclitax NAB are most stable.

A comparison of the ratio (as reported as a percentage) of thepercentage of albumin on the nanoparticles in the form of polymers (P)and oligomers (O) divided by the percentage of albumin on thenanoparticles in the form of monomers (M) (Table 25) is illustrated inthe bar graph of FIG. 24. Amongst all the compositions, thenab-paclitaxel sold under the trademark ABRAXANE® and Paclitax NAB aremost stable.

A comparison of the ratio (as reported as a percentage) of thepercentage of albumin on the nanoparticles in the form of polymers (P)and oligomers (O) divided by the percentage of albumin on thenanoparticles in the form of monomers (M) minus dimers (D) (Table 25) isillustrated in the bar graph of FIG. 25. Amongst all the compositions,the nab-paclitaxel sold under the trademark ABRAXANE® and Paclitax NABare most stable.

TABLE 26 Solubility (μg/ml) and dissolution kinetics of the compositionimmediately after reconstitution. Dissolution kinetics Sample Name N(number) Mean Margin of error Min Max comment ABRAXANE ® 11 66.1 5.353.5 82.2 Normal Paclitax NAB 1 63.3 — 63.3 63.3 Normal Albupax 1 40.0 —40.0 40.0 — PacliALL 3 85.0 21.2 76.0 93.0 Normal V5 FP 3 80.0 39.8 62.393.5 Normal V1 FP 3 66.2 22.4 58.0 75.9 Normal V2 FP 2 70.9 183.0 56.585.3 None V3 FP 3 74.8 13.6 68.8 79.5 Normal V4 FP 3 69.1 22.1 59.8 77.5Slower than normal

TABLE 27 Degree of sedimentation (stability) based on visual observationof the composition (1 indicates no sedimentation) immediately afterreconstitution. N Margin Sample Name (number) Mean of error Min MaxABRAXANE ® 17  1 0 1 1 Paclitax NAB 2 1 0 1 1 Albupax — — — — — PacliALL3 1 0 1 1 V5 FP 3 1 0 1 1 V1 FP 3 1 0 1 1 V2 FP 3 1 0 1 1 V3 FP 3 1 0 11 V4 FP 3 1 0 1 1 *The degree of sedimentation recited as: 1 - Novisible sedimentation (NVS); 2 - Streaming with NVS; 3 - Very slightsedimentation; 4 - Slight sedimentation; 5 - Sedimentation; 6 - Phaseseparation.

TABLE 28 Percentage of paclitaxel recovered after filtration through a0.2-μm syringe filter immediately after reconstitution. N Margin SampleName (number) Mean of error Min Max ABRAXANE ® 21 98.2 1.4 93.3 106.2Paclitax NAB 2 104.0 76.2 98 110 Albupax — — — — — PacliALL 5 43.0 24.127 72 V5 FP 3 95.7 8.7 92 99 V1 FP 3 92.3 18.0 84 97 V2 FP 2 92.0 63.587 97 V3 FP 3 97.0 7.5 94 100 V4 FP 3 99.0 13.1 95 105

Table 29 Solubility (μg/ml) and dissolution kinetics of the compositionafter storage of the reconstituted suspension for 24 hours at 40° C.Dissolution Sample N Margin Solubility kinetics Name (number) Mean oferror Min Max comment comment ABRAXANE ® 12 66.8 7.5 54.0 96.8 RegularNormal Paclitax NAB 1 44.9 — 44.9 44.9 Low solubility Normal Albupax — —— — — — Normal PacliALL 3 46.9 101.1 0.0 73.5 Insoluble/Regular — V5 FP2 74.1 2.5 73.9 74.3 ~Regular Normal V1 FP 2 0.0 0.0 0.0 0.0 InsolubleNone V2 FP 1 0.0 — 0.0 0.0 Insoluble None V3 FP 2 0.0 0.0 0.0 0.0Insoluble None V4 FP 2 0.0 0.0 0.0 0.0 Insoluble None

TABLE 30 Degree of sedimentation (stability) based on visual observation(1 indicates no sedimentation) and crystallinity (1 indicates presenceof crystalline paclitaxel) of the composition after storage of thereconstituted suspension for 24 hours at 40° C. Crystallinity SampleName N (number) Mean* Margin of error Min Max Crystallinity valueABRAXANE ® 17 1 0 1 1 No 0 Birefringence Paclitax NAB 2 1 0 1 1 No 0Birefringence Albupax 1 5 — 5 5 Birefringence 1 PacliALL 5 5 0 5 5Birefringence 1 V5 FP 2 5 0 5 5 Birefringence 1 V1 FP 2 5.5 6.4 5 6Birefringence 1 V2 FP 1 6 0 6 6 Birefringence 1 V3 FP 2 6 0 6 6Birefringence 1 V4 FP 2 6 0 6 6 Birefringence 1 *The degree ofsedimentation recited as: 1 - No visible sedimentation (NVS); 2 -Streaming with NVS; 3 - Very slight sedimentation; 4 - Slightsedimentation; 5 - Sedimentation; 6 - Phase separation.

TABLE 31 Degree of sedimentation (stability) based on visual observation(1 indicates no sedimentation) and crystallinity (1 indicates presenceof crystalline paclitaxel) of the composition after storage of thereconstituted suspension at 40° C. for 16 hours. Crystallinity SampleName Mean value V5 FP 2.7 0 V1 FP 5 1 V2 FP 5.5 1 V3 FP 5.3 0.66 V4 FP 50.66

TABLE 32 Percentage of paclitaxel recovered after filtration through a0.2-μm syringe filter afterstorage of the reconstituted suspension at40° C. for 16 hours. Sample Name Mean V5 FP 96.5 V1 FP 59.2 V2 FP 22.6V3 FP 50.1 V4 FP 36.2

TABLE 33 Degree of sedimentation (stability) based on visual observation(1 indicates no sedimentation) and crystallinity (1 indicates presenceof crystalline paclitaxel) of the composition after storage of thereconstituted suspension at 5° C. for 8 hours followed by storage at 25°C. for 8 hours. Crystallinity Sample Name Mean value V5 FP 1 0.333 V1 FP1.33 1 V2 FP 1.5 0 V3 FP 1.33 0 V4 FP 1 0

TABLE 34 Percentage of paclitaxel recovered after filtration through a0.2-μm syringe filter after storage of the reconstituted suspension at5° C. for 8 hours followed by storage at 25° C. for 8 hours. Sample NameMean V5 FP 98.7 V1 FP 72.7 V2 FP 64.0 V3 FP 91.0 V4 FP 92.0

TABLE 35 Percentage of paclitaxel recovered after filtration through a0.2-μm syringe filter after storage of the reconstituted suspension for24 hours at 40° C. N Margin Sample Name (number) Mean of error Min MaxABRAXANE ® 18 96.4 1.7 92.2 106.7 Paclitax NAB — — — — — Albupax 1 61 —61 61 PacliALL 1 69 — 69 69 V5 FP 2 86.3 36.8 83.4 89.2 V1 FP 2 13.5170.9 0.0 26.9 V2 FP 1 0.0 — 0.0 0.0 V3 FP 2 9.7 91.5 2.5 16.9 V4 FP 20.0 0.0 0.0 0.0

TABLE 36 D_(v4,3) (nm) measured after storage of the reconstitutedsuspension for 24 hours at 40° C. N Margin Sample Name (number) Mean oferror Min Max ABRAXANE ® 20 155.8 3.6 143.5 169.1 Paclitax NAB 2 118.448.9 114.5 122.2 Albupax 1 150.9 — 150.9 150.9 PacliALL 5 1052.5 3531.3231.5 2694 V5 FP 2 172.9 90.8 165.7 180 V1 FP 2 2556.5 7096.4 1998 3115V2 FP 1 2475.0 — 2475 2475 V3 FP 2 2408.0 5539.9 1972 2844 V4 FP 22431.0 7814.3 1816 3046

TABLE 37 Z average (nm) measured after storage of the reconstitutedsuspension for 24 hours at 40° C. N Margin Sample Name (number) Mean oferror Min Max ABRAXANE ® 20 147.7 2.2 139.5 155.8 Paclitax NAB 2 121.737.4 118.8 124.7 Albupax 1 157.7 — 157.7 157.7 PacliALL 5 1411.3 2634.2183.4 5098 V5 FP 2 156.9 60.3 152.2 161.7 V1 FP 2 2361.8 25414.2 361.74362 V2 FP 1 3435.0 — 3435 3435 V3 FP 2 692.0 4459.2 341.1 1043 V4 FP 25338.5 946.6 5264 5413

TABLE 38 Polydispersity index (PDI) measured after storage of thereconstituted suspension for 24 hours at 40° C. N Margin Sample Name(number) Mean of error Min Max ABRAXANE ® 20 0.11555 0.007141 0.0920.156 Paclitax NAB 2 0.1015  0.069884 0.096 0.107 Albupax 1 0.214  —0.214 0.214 PacliALL 5 0.3806  0.353505 0.178 0.862 V5 FP 2 0.141 0.1270619 0.131 0.151 V1 FP 2 0.6135  2.12829 0.446 0.781 V2 FP 1 0.315 — 0.315 0.315 V3 FP 2 0.8545  0.603542 0.807 0.902 V4 FP 2 0.368  1.43580.255 0.481

TABLE 39 DV5 (nm) measured after storage of the reconstituted suspensionfor 24 hours at 40° C. N Margin Sample Name (number) Mean of error MinMax ABRAXANE ® 20 76.2 2.4 65.2 84.7 Paclitax NAB 2 63.2 6.3 62.7 63.7Albupax 1 86.3 — 86.3 86.3 PacliALL 5 611.2 944.5 73.8 1760 V5 FP 2 73.894.0 66.4 81.2 V1 FP 2 834 8716.4 148 1520 V2 FP 1 1500.0 — 1500 1500 V3FP 2 172.5 209.6 156 189 V4 FP 2 1715.0 4637.7 1350 2080

TABLE 40 DV50 (nm) measured after storage of the reconstitutedsuspension for 24 hours at 40° C. Sample Name N (number) Mean Margin oferror Min Max ABRAXANE ® 20 140.2 4.138 124 154 Paclitax NAB 2 10738.11875 104 110 Albupax 1 142 — 142 142 PacliALL 5 1075.2 1466.9505 2002500 V5 FP 2 151 139.768 140 162 V1 FP 2 2795 10355.52 1980 3610 V2 FP 12337 — 2337 2337 V3 FP 2 2445 6035.44 1970 2920 V4 FP 2 2390 7369.591810 2970

TABLE 41 DV95 (nm) measured after storage of the reconstitutedsuspension for 24 hours at 40° C. Sample Name N (number) Mean Margin oferror Min Max ABRAXANE ® 20 290.9 7.695 267 336 Paclitax NAB 2 214.5133.41495 204 225 Albupax 1 249 — 249 249 PacliALL 5 3062.4 2951.1995496 5450 V5 FP 2 346 63.531 341 351 V1 FP 2 4305 21664 2600 6010 V2 FP 13917 — 3917 3917 V3 FP 2 4265 15565.1 3040 5490 V4 FP 2 3345 13023.822320 4370

TABLE 42 (DV90-DV10)/DV50 measured after storage of the reconstitutedsuspension for 24 hour at 40° C. N Margin Sample Name (number) Mean oferror Min Max ABRAXANE ® 20 1.20616  0.055545  1.03684 1.46048 PaclitaxNAB 2 1.08277  0.611737  1.03462 1.13091 Albupax 1 0.88803  — 0.888030.88803 PacliALL 5 3.76422  6.98718   0.736  13.8008   V5 FP 2 1.43079 2.077525  1.26728 1.59429 V1 FP 2 0.955875 6.498385  0.44444 1.46731 V2FP 1 0.82713  — 0.82713 0.82713 V3 FP 2 1.17052  6.101    0.690361.65068 V4 FP 2 0.517895 1.0347265 0.43646 0.59933

What is claimed is:
 1. A method of validating a commercial batch of apharmaceutical composition for medical use in a human individual,wherein the pharmaceutical composition comprises (a) nanoparticlescomprising paclitaxel coated with a coating comprising albumin and (b) anon-nanoparticle portion comprising albumin and paclitaxel, the methodcomprising: (1) obtaining a sample from the commercial batch; and (2)assessing suitability of the pharmaceutical composition for medical usein a human individual, comprising: separating the nanoparticles from thenon-nanoparticle portion, measuring a weight percentage of albumin inthe form of albumin polymers in the coating of the separatednanoparticles among the total albumin in the coating of the separatednanoparticles, and assessing the suitability of the pharmaceuticalcomposition for medical use in a human individual, wherein the weightpercentage of albumin in the form of albumin polymers in the coating ofthe separated nanoparticles among the total albumin in the coating ofthe separated nanoparticles being from about 15% to about 40% isindicative of suitability of the pharmaceutical composition for medicaluse; and (3) validating the commercial batch if the pharmaceuticalcomposition is suitable for medical use.
 2. The method of claim 1,wherein the method further comprises measuring a weight percentage ofalbumin in the form of albumin monomers in the coating of the separatednanoparticles among the total albumin in the coating of the separatednanoparticles, wherein the weight percentage of albumin in the form ofalbumin monomers in the coating of the separated nanoparticles among thetotal albumin in the coating of the separated nanoparticles being fromabout 40% to about 60% is indicative of suitability of thepharmaceutical composition for medical use.
 3. The method of claim 1,wherein the method further comprises measuring a weight percentage ofthe albumin in the coating of the separated nanoparticles compared tothe total weight of the separated nanoparticles, wherein the weightpercentage of the albumin in the coating of the separated nanoparticlescompared to the total weight of the separated nanoparticles being fromabout 15% to about 30% is indicative of suitability of thepharmaceutical composition for medical use.
 4. The method of claim 1,further comprising determining a weight ratio of albumin in the coatingof the separated nanoparticles to paclitaxel in the nanoparticles,wherein the weight ratio of albumin in the coating of the separatednanoparticles to paclitaxel in the nanoparticles being from about 1:2 toabout 1:6 in the nanoparticles is indicative of suitability of thepharmaceutical composition for medical use.
 5. The method of claim 1,further comprising determining a thickness of the coating of thenanoparticles by cryogenic transmission electron microscopy, wherein thethickness being about 5-7 nm as determined by cryogenic transmissionelectron microscopy is indicative of suitability of the pharmaceuticalcomposition for medical use.
 6. The method of claim 1, furthercomprising determining a solubility of the nanoparticles in a 5% albuminsolution by dynamic light scattering, wherein the solubility of thenanoparticles being from about 50 μg/ml to about 100 μg/ml in the 5%human albumin solution is indicative of suitability of thepharmaceutical composition for medical use.
 7. The method of claim 1,further comprising determining a paclitaxel crystallinity of thepaclitaxel in the nanoparticles of the pharmaceutical composition byX-ray diffraction or polarized light microscopy, wherein anon-crystalline state of the paclitaxel is indicative of suitability ofthe pharmaceutical composition for medical use.
 8. The method of claim1, further comprising determining a paclitaxel recovery following a 0.2micron filtration of the pharmaceutical composition, wherein thepaclitaxel recovery being at least 80% is indicative of suitability ofthe pharmaceutical composition for medical use.
 9. The method of claim6, wherein the determination of solubility is carried out after storage.10. The method of claim 1, further comprising measuring a weightpercentage of albumin in the form of albumin dimers in the coating ofthe separated nanoparticles among the total albumin in the coating ofthe separated nanoparticles, wherein the weight percentage of albumin inthe form of albumin dimers in the coating of the separated nanoparticlesamong the total albumin in the coating of the separated nanoparticlesbeing from about 15% to about 30% is indicative of suitability of thepharmaceutical composition for medical use.
 11. The method of claim 1,further comprising measuring a weight percentage of albumin in the formof albumin oligomers in the coating of the separated nanoparticles amongthe total albumin in the coating of the separated nanoparticles, whereinthe weight percentage of albumin in the form of albumin oligomers in thecoating of the separated nanoparticles among the total albumin in thecoating of the separated nanoparticles being from about 7% to about 15%is indicative of suitability of the pharmaceutical composition formedical use.
 12. The method of claim 1, further comprising determining aparticle size of the nanoparticles using dynamic light scattering,wherein the particle size of the nanoparticles being less than 200 nm isindicative of suitability of the pharmaceutical composition for medicaluse.
 13. The method of claim 1, further comprising determining apolydispersity index of the nanoparticles in the pharmaceuticalcomposition using dynamic light scattering, wherein the polydispersityindex of the nanoparticles being less than 0.3 is indicative ofsuitability of the pharmaceutical composition for medical use.
 14. Themethod of claim 1, further comprising determining a span of sizedistribution of the nanoparticles in the pharmaceutical composition as(D_(v90)−D_(v10))/D_(v50) using dynamic light scattering, whereinD_(v90) is the particle diameter where 90% of the volume of allnanoparticles is contained in nanoparticles with smaller diameters;D_(v10) is the particle diameter where 10% of the volume of allnanoparticles is contained in nanoparticles with smaller diameters; andD_(v50) is the volume-weighted median particle diameter; and wherein thespan of size distribution being from about 0.8 to about 1.5 isindicative of suitability of the pharmaceutical composition for medicaluse.
 15. The method of claim 1, further comprising determining a surfacepotential of the nanoparticles using microelectrophoresis,electrophoretic light scattering, dynamic electrophoretic mobility, ortunable resistive pulse sensing, wherein the surface potential of thenanoparticles being from about −20 mV to about −35 mV is indicative ofsuitability of the nanoparticles for medical use.
 16. The method ofclaim 1, further comprising measuring a weight percentage of thepaclitaxel in the separated nanoparticles among the total paclitaxel inthe pharmaceutical composition using reversed-phase high performanceliquid chromatography, wherein the weight percentage of the paclitaxelin the separated nanoparticles among the total paclitaxel in thepharmaceutical composition being at least 95% is indicative ofsuitability of the pharmaceutical composition for medical use.
 17. Themethod of claim 1, further comprising measuring a weight percentage ofthe albumin that is in the non-nanoparticle portion among the totalalbumin in the pharmaceutical composition using size-exclusionchromatography, wherein the weight percentage of the albumin that is inthe non-nanoparticle portion among the total albumin in thepharmaceutical composition being at least 95% is indicative ofsuitability of the pharmaceutical composition for medical use.
 18. Themethod of claim 1, wherein the pharmaceutical composition has a weightratio of total albumin to total paclitaxel in the range from about 3:1to about 7.9:1 or from about 10:1 to about 17:1.
 19. The method of claim1, wherein the albumin is human albumin.
 20. The method of claim 1,wherein the average diameter of the nanoparticles in the pharmaceuticalcomposition is less than about 200 nm as determined by dynamic lightscattering.
 21. A commercial batch of a pharmaceutical composition formedical use in a human individual, wherein the pharmaceuticalcomposition comprises (a) nanoparticles comprising paclitaxel coatedwith a coating comprising albumin and (b) a non-nanoparticle portioncomprising albumin and paclitaxel, and wherein the commercial batch isvalidated by method of claim
 1. 22. The method of claim 1, furthercomprising measuring a weight percentage of albumin in the form ofalbumin monomers in the coating of the separated nanoparticles among thetotal albumin in the coating of the separated nanoparticles, wherein theweight percentage of albumin in the form of albumin monomers in thecoating of the separated nanoparticles among the total albumin in thecoating of the separated nanoparticles being less than about 51% isindicative of the pharmaceutical composition for medical use.
 23. Themethod of claim 1, further comprising measuring a weight percentage ofalbumin in the form of albumin monomers, albumin oligomers, and albuminpolymers in the coating of the separated nanoparticles among the totalalbumin in the coating of the separated nanoparticles, wherein a ratioof the sum of the weight percentage of albumin in the form of albuminpolymers in the coating of the separated nanoparticles and the weightpercentage of the albumin in the form of albumin oligomers in thecoating of the separated nanoparticles compared to the weight percentageof albumin in the form of albumin monomers in the coating of theseparated nanoparticles being more than about 65% is indicative of thepharmaceutical composition for medical use.
 24. A method of releasing acommercial batch of a pharmaceutical composition comprising (a)nanoparticles comprising paclitaxel coated with a coating comprisingalbumin and (b) a non-nanoparticle portion comprising albumin andpaclitaxel, the method comprising: (1) assessing suitability of thepharmaceutical composition for medical use in a human individual,comprising: separating the nanoparticles from the non-nanoparticleportion, measuring a weight percentage of albumin in the form of albuminpolymers in the coating of the separated nanoparticles among the totalalbumin in the coating of the separated nanoparticles, and assessing thesuitability of the pharmaceutical composition for medical use in a humanindividual, wherein the weight percentage of albumin in the form ofalbumin polymers in the coating of the separated nanoparticles among thetotal albumin in the coating of the separated nanoparticles being fromabout 15% to about 40% is indicative of suitability of thepharmaceutical composition for medical use; and (2) releasing thecommercial batch if the pharmaceutical composition is suitable formedical use.
 25. A method of processing a sample of a pharmaceuticalcomposition to assess suitability of the pharmaceutical composition formedical use in a human individual, the pharmaceutical compositioncomprising (a) nanoparticles comprising paclitaxel coated with a coatingcomprising albumin and (b) a non-nanoparticle portion comprising albuminand paclitaxel, the method comprising: (1) obtaining the sample from acommercial batch; and (2) processing the sample to assess suitability ofthe pharmaceutical composition for medical use in a human individual,comprising: separating the nanoparticles from the non-nanoparticleportion, measuring a weight percentage of albumin in the form of albuminpolymers in the coating of the separated nanoparticles among the totalalbumin in the coating of the separated nanoparticles, and assessing thesuitability of the pharmaceutical composition for medical use in a humanindividual, wherein the weight percentage of albumin in the form ofalbumin polymers in the coating of the separated nanoparticles among thetotal albumin in the coating of the separated nanoparticles being fromabout 15% to about 40% is indicative of suitability of thepharmaceutical composition for medical use.